47 results on '"De Felice, M."'
Search Results
2. Regulation of Isoleucine and Valine Biosynthesis
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Maurizio Iaccarino, John Guardiola, Renée Favre, and De Felice M
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chemistry.chemical_classification ,Acetolactate synthase ,Acetolactate synthase activity ,Transamination ,Stereochemistry ,Threonine Dehydratase ,Biology ,chemistry ,Biochemistry ,Valine ,biology.protein ,Isoleucine ,Threonine ,Amino acid synthesis - Abstract
Publisher Summary This chapter discusses the regulation of isoleucine and valine biosynthesis. Isoleucine and valine biosynthesis is a model system for the study of mechanisms by which the cell coordinates different parts of its total metabolism. Threonine deaminase is the first enzyme in isoleucine biosynthesis. A different threonine deaminase, termed catabolic, appears only in anaerobiosis and in the absence of glucose. A third threonine deaminase, present in Escherichia coli K-12 and Salmonella Typhimurium, deaminates only D-threonine, and its presence explains why ilvA mutants of these organisms grow upon addition of D-threonine to the medium. The second step in the pathway is common to isoleucine and valine biosynthesis and is catalyzed by acetolactate synthase activity. It involves the conversion of either two molecules of pyruvate to form α-acetolactate, or one molecule of pyruvate and one molecule of α-ketobutyrate to form α-aceto-α-hydroxybutyrate. The acetolactate synthase isoenzymes have some relation to lysine biosynthesis. The third step in isoleucine biosynthesis and the second in valine biosynthesis are catalyzed by the enzyme isomeroreductase. The last intermediates in the pathway for isoleucine and valine biosynthesis are the keto acids from which the corresponding amino acids are synthesized by transamination.
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- 1978
3. Identification and characterization of the proBA operon of Streptococcus bovis
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A L Basso, L Ferrara, Margherita Sacco, Ezio Ricca, C Campanile, M De Felice, Giuseppe Forlani, Rosangela Marasco, Campanile, C, Forlani, G, Basso, A, Marasco, Rosangela, Ricca, E, Sacco, Margherita, Ferrara, L. AND DE FELICE M., Basso, Al, Marasco, R, Ricca, Ezio, Sacco, M, Ferrara, L, DE FELICE, M., and DE FELICE, Maurilio
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DNA, Bacterial ,Enzyme complex ,Proline ,Operon ,DNA, Recombinant ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,Plasmid ,Escherichia coli ,medicine ,Genomic library ,Gene Library ,Genetics ,Ecology ,biology ,Phosphotransferases ,Streptococcus bovis ,biology.organism_classification ,Complementation ,genomic DNA ,Biochemistry ,Oxidoreductases ,Research Article ,Food Science ,Biotechnology - Abstract
A genomic DNA library of the rumen bacterium Streptococcus bovis was constructed in Escherichia coli, and recombinant plasmids able to complement proA and proB mutations of the host were found. Southern hybridization and restriction analysis showed that a 3.5-kb fragment of S. bovis DNA contained two genes, organized in an operon and coding for enzymes functionally similar to the glutamyl phosphate reductase-glutamyl kinase enzyme complex that in E. coli catalyzes the first step of proline biosynthesis. Complementation of the E. coli mutations was observed with the fragment inserted in both orientations, which suggested that the S. bovis proBA operon was transcribed from its own promoter. Genetic and biochemical data suggested that the proline biosynthetic pathway of S. bovis is similar to the one previously characterized for E. coli.
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- 1993
4. Surface display of recombinant proteins on Bacillus subtilis spores
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Rachele Isticato, Giuseppina Cangiano, Donata Medaglini, Annalisa Ciabattini, H. T. Tran, Ezio Ricca, M De Felice, Gianni Pozzi, Marco R. Oggioni, Isticato R, Cangiano G, Tran HT, Ciabattini A, Medaglini D, Oggioni MR, De Felice M, Pozzi G, Ricca E, Isticato, Rachele, Cangiano, Giuseppina, Tran, Ht, Ciabattini, A, Medaglini, D, Oggioni, Mr, DE FELICE, Maurilio, Pozzi, G, Ricca, Ezio, Isticato, R, DE FELICE, M, Ricca, E., Isticato, R., Cangiano, G., Tran, H. T., Ciabattini, A., Medaglini, D., Oggioni, M. R., and Pozzi, G.
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Recombinant Fusion Proteins ,Molecular Sequence Data ,Heterologous ,Dot blot ,Cell Surfaces ,Bacillus subtilis ,delivery system ,medicine.disease_cause ,Microbiology ,Endospore ,law.invention ,Mice ,Bacterial Proteins ,law ,vaccine ,Tetanus Toxoid ,medicine ,Animals ,Amino Acid Sequence ,Molecular Biology ,Spores, Bacterial ,biology ,Toxin ,fungi ,Membrane Proteins ,Cell sorting ,Flow Cytometry ,biology.organism_classification ,Spore ,Mutagenesis, Insertional ,Immunoglobulin G ,Recombinant DNA ,Bacillus subtilis spores - Abstract
We developed a novel surface display system based on the use of bacterial spores. A protein of the Bacillus subtilis spore coat, CotB, was found to be located on the spore surface and used as fusion partner to express the 459-amino-acid C-terminal fragment of the tetanus toxin (TTFC). Western, dot blot and fluorescent-activated cell sorting analyses were used to monitor TTFC surface expression on purified spores. We estimated that more than 1.5 × 10 3 TTFC molecules were exposed on the surface of each spore and recognized by TTFC-specific antibodies. The efficient surface presentation of the heterologous protein, together with the simple purification procedure and the high stability and safety record of B. subtilis spores, makes this spore-based display system a potentially powerful approach for surface expression of bioactive molecules.
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- 2001
5. On the fate of ingested Bacillus spores
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Maria R Spinosa, Marco R. Oggioni, Ezio Ricca, Maurilio De Felice, Lorenzo Morelli, Tiziana Braccini, Gianni Pozzi, Spinosa MR, Braccini T, Ricca E, De Felice M, Morelli L, Pozzi G, Oggioni MR, Spinosa, Mr, Braccini, T, Ricca, Ezio, DE FELICE, M, Morelli, L, Pozzi, G, Oggioni, Mr, Spinosa, M. R., Braccini, T., Ricca, E., DE FELICE, Maurilio, Morelli, L., Pozzi, G., and Oggioni, M. R.
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Bacillus ,Administration, Oral ,Bacillus subtilis ,Microbial Sensitivity Tests ,Microbiology ,Bile Acids and Salts ,chemistry.chemical_compound ,Mice ,Animals ,Molecular Biology ,Spores, Bacterial ,Mice, Inbred BALB C ,Taurodeoxycholic Acid ,biology ,Probiotics ,Bacillus clausii ,fungi ,General Medicine ,biology.organism_classification ,Bacillales ,Spore ,Intestines ,Cereus ,chemistry ,Germination ,Female ,Lymph Nodes ,Taurodeoxycholic acid ,probiotic ,Deoxycholic Acid - Abstract
Spores of various Bacillus species, including B. subtilis, B. cereus and B. clausii, are used as probiotics, although they are generally absent from the normal microflora of man. We used two nonpathogenic Bacillus species, B. subtilis and B. clausii, to follow the fate of spores inoculated intragastrically in mice. We did not find detectable amounts of vegetative cells in intestinal samples, probably because of high toxicity of the conjugated bile salt taurodeoxycholic acid against Bacillus species. Both spores and cells were detected in the lymph nodes and spleen of one mouse. Our results indicate that Bacillus is present in the intestinal tract solely as spores and that nonpathogenic Bacillus spores may germinate in lymphoid organs, a finding reminiscent of B. anthracis germination in macrophages. These results indicate that any claimed probiotic effect of B. subtilis should be due to spores or, alternatively, to vegetative growth outside the intestine.
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- 2000
6. A mouse model for hereditary thyroid dysgenesis and cleft palate
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Hans R. Schöler, Roberto Di Lauro, Claudio Arra, Konstantinos Anastassiadis, Elio Biffali, Vincenzo Macchia, Angela Mariano, Catherine E. Ovitt, Alina Rodriguez-Mallon, Marie Genevieve Mattei, Paolo Emidio Macchia, Mario De Felice, DE FELICE, M, Ovitt, C, Biffali, E, RODRIGUEZ MALLON, A, Arra, C, Anastassiadis, K, Macchia, PAOLO EMIDIO, Mattei, Mg, Mariano, A, Scholer, H, Macchia, Vincenzo, DI LAURO, R., De Felice, M., Ovitt, C., Biffali, E., Rodriguez-Mallon, A., Arra, C., Anastassiadis, K., Macchia, P. E., Mattei, M. -G., Mariano, A., Scholer, H., Macchia, V., and Di Lauro, R.
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endocrine system ,medicine.medical_specialty ,endocrine system diseases ,Transcription Factor ,DNA-Binding Protein ,Morphogenesis ,Thyroid Gland ,Ectoderm ,Biology ,Thyroid dysgenesis ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Hypothyroidism ,Internal medicine ,Genetics ,medicine ,Morphogenesi ,Animals ,030304 developmental biology ,Mice, Knockout ,0303 health sciences ,Animal ,Thyroid ,Endoderm ,Foregut ,Forkhead Transcription Factors ,Forkhead Transcription Factor ,Repressor Protein ,medicine.disease ,Congenital hypothyroidism ,Cleft Palate ,DNA-Binding Proteins ,Repressor Proteins ,Disease Models, Animal ,medicine.anatomical_structure ,Chromosome 4 ,Endocrinology ,030220 oncology & carcinogenesis ,FOXE1 ,Transcription Factors - Abstract
Alteration of thyroid gland morphogenesis (thyroid dysgenesis) is a frequent human malformation. Among the one in three to four thousand newborns in which congenital hypothyroidism is detected, 80% have either an ectopic, small and sublingual thyroid, or have no thyroid tissue1. Most of these cases appear sporadically, although a few cases of recurring familial thyroid dysgenesis have been described2. The lack of evidence for hereditary thyroid dysgenesis may be due to the severity of the hypothyroid phenotype. Neonatal screening and early thyroid hormone therapy have eliminated most of the clinical consequences of hypothyroidism such that the heritability of this condition may become apparent in the near future. We have recently cloned cDNA encoding a forkhead domain-containing transcription factor, TTF-2, and have located the position of the gene, designated Titf2, to mouse chromosome 4 (ref. 3). Titf2 is expressed in the developing thyroid, in most of the foregut endoderm and in craniopharyngeal ectoderm, including Rathke's pouch3. Expression of Titf2 in thyroid cell precursors is down-regulated as they cease migration, suggesting that this factor is involved in the process of thyroid gland morphogenesis. Here we show that Titf2-null mutant mice exhibit cleft palate and either a sublingual or completely absent thyroid gland. Thus, mutation of Titf2 –/– results in neonatal hypothyroidism that shows similarity to thyroid dysgenesis in humans.
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- 1998
7. Induction of UCP2 mRNA by thyroid hormones in rat heart
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Maria Moreno, Antonia Lanni, Daniel Ricquier, Christophe Fleury, Fernando Goglia, Assunta Lombardi, Mario De Felice, Lanni, Antonia, DE FELICE, M, Lombardi, A, Moreno, M, Fleury, C, Ricqier, D, Goglia, F., A., Lanni, DE FELICE, Mario, A., Lombardi, M., Moreno, C., Fleury, D., Ricquier, F., Goglia, Lanni, A., De Felice, M., and Lombardi, Assunta
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Male ,medicine.medical_specialty ,Transcription, Genetic ,Thyroid Gland ,Biophysics ,Skeletal muscle ,Biology ,Kidney ,Hyperthyroidism ,Iodide Peroxidase ,Biochemistry ,Ion Channels ,Iopanoic acid ,Mitochondrial Proteins ,Hypothyroidism ,Structural Biology ,Internal medicine ,Brown adipose tissue ,Uncoupling protein ,Genetics ,medicine ,Animals ,Uncoupling Protein 2 ,RNA, Messenger ,Rats, Wistar ,Muscle, Skeletal ,Molecular Biology ,Triiodothyronine ,Myocardium ,Membrane Transport Proteins ,Heart ,Cell Biology ,Thermogenin ,Rats ,Thyroid hormone ,Thyroxine ,medicine.anatomical_structure ,Endocrinology ,Liver ,Organ Specificity ,Protein Biosynthesis ,Propylthiouracil ,medicine.drug - Abstract
The possible regulation of the expression of uncoupling protein-2 (UCP2) mRNA by thyroid hormones in different tissues was examined in rats, Triiodothyronine (T-3) was found to produce an organ-specific enhancement of UCP2 expression in rat tissues. The effect of T-3 was markedly observed in heart, whereas a moderate effect was seen in skeletal muscle and no effect in kidney or liver, These results suggest that UCP2 is a protein that may be involved in the nuclear-mediated effect of T-3 on resting metabolic rate in the rat. (C) 1997 Federation of European Biochemical Societies.
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- 1997
8. In vivo footprinting analysis of Lrp binding to the ilvIH promoter region of Escherichia coli
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Rosangela Marasco, Margherita Sacco, M De Felice, F La Cara, Mario Varcamonti, Ezio Ricca, Marasco, R, Varcamonti, Mario, LA CARA, F, Ricca, E, DE FELICE, M. AND SACCO M., Marasco, Rosangela, Varcamonti, M, Sacco, Margherita, Ricca, Ezio, DE FELICE, M, Sacco, M., and DE FELICE, Maurilio
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DNA, Bacterial ,Operon ,Molecular Sequence Data ,Regulatory Sequences, Nucleic Acid ,Methylation ,Microbiology ,DNA-binding protein ,Bacterial Proteins ,Leucine ,Transcription (biology) ,Escherichia coli ,Leucine-responsive regulatory protein ,Binding site ,Promoter Regions, Genetic ,Molecular Biology ,Transcription factor ,Binding Sites ,Base Sequence ,biology ,Escherichia coli Proteins ,Promoter ,Leucine-Responsive Regulatory Protein ,Molecular biology ,Footprinting ,DNA-Binding Proteins ,Kinetics ,biology.protein ,lipids (amino acids, peptides, and proteins) ,Transcription Factors ,Research Article - Abstract
An in vivo footprinting analysis of the ilvIH regulatory region of Escherichia coli showed that the transcription activator Lrp binds to six sites, scattered over 250 bp upstream of the transcriptional start point. When Lrp-mediated activation was impaired by the presence of exogenous leucine, only one promoter-distal site (site 2) was partially protected by Lrp binding. Equilibrium dialysis experiments showed the formation of an Lrp-leucine complex in vitro. These results suggest that leucine negatively affects ilvIH transcription because its interaction with Lrp reduces the efficiency of binding of the regulatory protein to the promoter region.
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- 1994
9. Retinoic Acid Induces Embryonic Stem Cells (ESCs) Transition to 2 Cell-Like State Through a Coordinated Expression of Dux and Duxbl1
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Daniela Tagliaferri, Pellegrino Mazzone, Teresa M. R. Noviello, Martina Addeo, Tiziana Angrisano, Luigi Del Vecchio, Feliciano Visconte, Vitalba Ruggieri, Sabino Russi, Antonella Caivano, Irene Cantone, Mario De Felice, Michele Ceccarelli, Luigi Cerulo, Geppino Falco, Tagliaferri, D., Mazzone, P., Noviello, T. M. R., Addeo, M., Angrisano, T., Del Vecchio, L., Visconte, F., Ruggieri, V., Russi, S., Caivano, A., Cantone, I., De Felice, M., Ceccarelli, M., Cerulo, L., and Falco, G.
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0301 basic medicine ,Cell ,Population ,Retinoic acid ,ESC ,Biology ,03 medical and health sciences ,chemistry.chemical_compound ,Cell and Developmental Biology ,0302 clinical medicine ,metastate ,medicine ,retinoic acid ,Inner cell mass ,Blastocyst ,education ,lcsh:QH301-705.5 ,reproductive and urinary physiology ,Original Research ,education.field_of_study ,urogenital system ,2-cell like ,ESCs ,pluripotency ,Cell Biology ,Embryonic stem cell ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,lcsh:Biology (General) ,030220 oncology & carcinogenesis ,embryonic structures ,Maternal to zygotic transition ,biological phenomena, cell phenomena, and immunity ,Reprogramming ,Developmental Biology - Abstract
Embryonic stem cells (ESCs) are derived from inner cell mass (ICM) of the blastocyst. In serum/LIF culture condition, they show variable expression of pluripotency genes that mark cell fluctuation between pluripotency and differentiation metastate. The ESCs subpopulation marked by zygotic genome activation gene (ZGA) signature, including Zscan4, retains a wider differentiation potency than epiblast-derived ESCs. We have recently shown that retinoic acid (RA) significantly enhances Zscan4 cell population. However, it remains unexplored how RA initiates the ESCs to 2-cell like reprogramming. Here we found that RA is decisive for ESCs to 2C-like cell transition, and reconstructed the gene network surrounding Zscan4. We revealed that RA regulates 2C-like population co-activating Dux and Duxbl1. We provided novel evidence that RA dependent ESCs to 2C-like cell transition is regulated by Dux, and antagonized by Duxbl1. Our suggested mechanism could shed light on the role of RA on ESC reprogramming.
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- 2020
10. Insight into nephrocan function in mouse endoderm patterning
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Valeria Lucci, Federica Amodio, Elena Amendola, Mario De Felice, Maria De Angelis, Nicola Antonino Russo, Luca Roberto, Filomena Russo, Pina Marotta, Silvia Buonaiuto, Ilaria Guerriero, Geppino Falco, Anna Iervolino, Antonio Marino, Feliciano Visconte, Martina Addeo, Addeo, M., Buonaiuto, S., Guerriero, I., Amendola, E., Visconte, F., Marino, A., De Angelis, M. T., Russo, F., Roberto, L., Marotta, P., Antonino Russo, N., Iervolino, A., Amodio, F., De Felice, M., Lucci, V., and Falco, G.
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0301 basic medicine ,Nephrocan gene ,Mice ,0302 clinical medicine ,Intercellular Signaling Peptides and Protein ,CRISPR ,Protein Isoforms ,Spectroscopy ,Gene Editing ,Mice, Knockout ,differentiation, definitive endoderm ,Endoderm ,Gene Expression Regulation, Developmental ,Cell Differentiation ,General Medicine ,embryonic stem cells ,Phenotype ,Computer Science Applications ,Cell biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Differentiation ,embryonic structures ,Gene Targeting ,Intercellular Signaling Peptides and Proteins ,Transcriptional variant ,(CRISPR)/CRISPR-associated systems 9 (Cas9) ,animal structures ,Germ layer ,[object Object] ,Biology ,Catalysis ,Article ,Mouse model ,Inorganic Chemistry ,03 medical and health sciences ,medicine ,Definitive endoderm ,Animals ,Physical and Theoretical Chemistry ,Molecular Biology ,Gene ,Body Patterning ,transcriptional variants ,Animal ,Organic Chemistry ,Alternative splicing ,Embryonic stem cell ,030104 developmental biology ,Genetic Loci ,Function (biology) - Abstract
Endoderm-derived organs as liver and pancreas are potential targets for regenerative therapies, and thus, there is great interest in understanding the pathways that regulate the induction and specification of this germ layer. Currently, the knowledge of molecular mechanisms that guide the in vivo endoderm specification is restricted by the lack of early endoderm specific markers. Nephrocan (Nepn) is a gene whose expression characterizes the early stages of murine endoderm specification (E7.5&ndash, 11.5) and encodes a secreted N-glycosylated protein. In the present study, we report the identification of a new transcript variant that is generated through alternative splicing. The new variant was found to have differential and tissue specific expression in the adult mouse. In order to better understand Nepn role during endoderm specification, we generated Nepn knock-out (KO) mice. Nepn&minus, /&minus, mice were born at Mendelian ratios and displayed no evident phenotype compared to WT mice. In addition, we produced nullizygous mouse embryonic stem cell (mESC) line lacking Nepn by applying (CRISPR)/CRISPR-associated systems 9 (Cas9) and employed a differentiation protocol toward endoderm lineage. Our in vitro results revealed that Nepn loss affects the endoderm differentiation impairing the expression of posterior foregut-associated markers.
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- 2020
11. FOXE1-Dependent Regulation of Macrophage Chemotaxis by Thyroid Cells In Vitro and In Vivo
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Irene Cantone, Gabriella De Vita, Matteo Esposito, Marta De Menna, Mario De Felice, Carmen Moccia, Sara C Credendino, Luigi Di Guida, Credendino, S. C., De Menna, M., Cantone, I., Moccia, C., Esposito, M., Di Guida, L., De Felice, M., and De Vita, G.
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0301 basic medicine ,Chemokine ,Macrophage ,Thyroid Gland ,chemokines ,Transcriptome ,Mice ,0302 clinical medicine ,Biology (General) ,Thyroid cancer ,Cells, Cultured ,Spectroscopy ,Mice, Knockout ,biology ,Thyroid ,Chemotaxi ,General Medicine ,M2 Macrophage ,Computer Science Applications ,Chemistry ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,tumour microenvironment ,Human ,QH301-705.5 ,TAMs ,Article ,Catalysis ,Macrophage chemotaxis ,Inorganic Chemistry ,03 medical and health sciences ,medicine ,Thyroid Neoplasms ,Physical and Theoretical Chemistry ,QD1-999 ,Molecular Biology ,Animal ,In Vitro Technique ,Organic Chemistry ,Chemotaxis ,Forkhead Transcription Factor ,medicine.disease ,Rats, Inbred F344 ,Disease Models, Animal ,030104 developmental biology ,TAM ,biology.protein ,Cancer research ,Rat ,FOXE1 - Abstract
Forkhead box E1 (FOXE1) is a lineage-restricted transcription factor involved in thyroid cancer susceptibility. Cancer-associated polymorphisms map in regulatory regions, thus affecting the extent of gene expression. We have recently shown that genetic reduction of FOXE1 dosage modifies multiple thyroid cancer phenotypes. To identify relevant effectors playing roles in thyroid cancer development, here we analyse FOXE1-induced transcriptional alterations in thyroid cells that do not express endogenous FOXE1. Expression of FOXE1 elicits cell migration, while transcriptome analysis reveals that several immune cells-related categories are highly enriched in differentially expressed genes, including several upregulated chemokines involved in macrophage recruitment. Accordingly, FOXE1-expressing cells induce chemotaxis of co-cultured monocytes. We then asked if FOXE1 was able to regulate macrophage infiltration in thyroid cancers in vivo by using a mouse model of cancer, either wild type or with only one functional FOXE1 allele. Expression of the same set of chemokines directly correlates with FOXE1 dosage, and pro-tumourigenic M2 macrophage infiltration is decreased in tumours with reduced FOXE1. These data establish a novel link between FOXE1 and macrophages recruitment in the thyroid cancer microenvironment, highlighting an unsuspected function of this gene in the crosstalk between neoplastic and immune cells that shape tumour development and progression.
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- 2021
12. FOXE1 Gene Dosage Affects Thyroid Cancer Histology and Differentiation In Vivo
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Carmen Moccia, Elena Amendola, Eduardo Clery, Sara C Credendino, Giuliana D'Avino, Gabriella De Vita, Arturo Brunetti, Luigi Di Guida, Adelaide Greco, Mario De Felice, Claudio Bellevicine, Credendino, S. C., Moccia, C., Amendola, E., D'Avino, G., Di Guida, L., Clery, E., Greco, A., Bellevicine, C., Brunetti, A., De Felice, M., and De Vita, G.
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0301 basic medicine ,mouse model ,proliferation ,Biology ,Article ,Catalysis ,Papillary thyroid cancer ,lcsh:Chemistry ,thyroid cancer susceptibility ,Inorganic Chemistry ,Thyroid carcinoma ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Neoplastic transformation ,Physical and Theoretical Chemistry ,lcsh:QH301-705.5 ,Molecular Biology ,Thyroid cancer ,Spectroscopy ,Oncogene ,Organic Chemistry ,Thyroid ,Cancer ,differentiation ,General Medicine ,FOXE1 ,medicine.disease ,Computer Science Applications ,030104 developmental biology ,medicine.anatomical_structure ,lcsh:Biology (General) ,lcsh:QD1-999 ,030220 oncology & carcinogenesis ,Cancer research - Abstract
The transcription factor Forkhead box E1 (FOXE1) is a key player in thyroid development and function and has been identified by genome-wide association studies as a susceptibility gene for papillary thyroid cancer. Several cancer-associated polymorphisms fall into gene regulatory regions and are likely to affect FOXE1 expression levels. However, the possibility that changes in FOXE1 expression modulate thyroid cancer development has not been investigated. Here, we describe the effects of FOXE1 gene dosage reduction on cancer phenotype in vivo. Mice heterozygous for FOXE1 null allele (FOXE1+/&minus, ) were crossed with a BRAFV600E-inducible cancer model to develop thyroid cancer in either a FOXE1+/+ or FOXE1+/&minus, genetic background. In FOXE1+/+ mice, cancer histological features are quite similar to that of human high-grade papillary thyroid carcinomas, while cancers developed with reduced FOXE1 gene dosage maintain morphological features resembling less malignant thyroid cancers, showing reduced proliferation index and increased apoptosis as well. Such cancers, however, appear severely undifferentiated, indicating that FOXE1 levels affect thyroid differentiation during neoplastic transformation. These results show that FOXE1 dosage exerts pleiotropic effects on thyroid cancer phenotype by affecting histology and regulating key markers of tumor differentiation and progression, thus suggesting the possibility that FOXE1 could behave as lineage-specific oncogene in follicular cell-derived thyroid cancer.
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- 2020
13. DNAJC17 is localized in nuclear speckles and interacts with splicing machinery components
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Elena Amendola, Andrea Scaloni, G. Ferrandino, C. Sette, A. Pascarella, G De Vita, Sara Carmela Credendino, M De Felice, O. Spadaro, P. Bielli, Michele Ceccarelli, Chiara D'Ambrosio, B. Miranda, Carmen Moccia, Fulvio D'Angelo, Pascarella, A., Ferrandino, G., Credendino, S. C., Moccia, C., D'Angelo, F., Miranda, B., D'Ambrosio, C., Bielli, P., Spadaro, O., Ceccarelli, M., Scaloni, A., Sette, C., De Felice, M., De Vita, G., and Amendola, E.
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0301 basic medicine ,Proteomics ,lcsh:Medicine ,Biology ,Interactome ,Article ,Transcriptome ,03 medical and health sciences ,Protein Interaction Mapping ,Humans ,lcsh:Science ,Gene ,Cell Nucleus ,Gene knockdown ,Settore BIO/16 ,Multidisciplinary ,Alternative Splicing ,HSP40 Heat-Shock Proteins ,HeLa Cells ,Spliceosomes ,Alternative splicing ,lcsh:R ,Cell biology ,030104 developmental biology ,RNA splicing ,Knockout mouse ,lcsh:Q ,Minigene - Abstract
DNAJC17 is a heat shock protein (HSP40) family member, identified in mouse as susceptibility gene for congenital hypothyroidism. DNAJC17 knockout mouse embryos die prior to implantation. In humans, germline homozygous mutations in DNAJC17 have been found in syndromic retinal dystrophy patients, while heterozygous mutations represent candidate pathogenic events for myeloproliferative disorders. Despite widespread expression and involvement in human diseases, DNAJC17 function is still poorly understood. Herein, we have investigated its function through high-throughput transcriptomic and proteomic approaches. DNAJC17-depleted cells transcriptome highlighted genes involved in general functional categories, mainly related to gene expression. Conversely, DNAJC17 interactome can be classified in very specific functional networks, with the most enriched one including proteins involved in splicing. Furthermore, several splicing-related interactors, were independently validated by co-immunoprecipitation and in vivo co-localization. Accordingly, co-localization of DNAJC17 with SC35, a marker of nuclear speckles, further supported its interaction with spliceosomal components. Lastly, DNAJC17 up-regulation enhanced splicing efficiency of minigene reporter in live cells, while its knockdown induced perturbations of splicing efficiency at whole genome level, as demonstrated by specific analysis of RNAseq data. In conclusion, our study strongly suggests a role of DNAJC17 in splicing-related processes and provides support to its recognized essential function in early development.
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- 2018
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14. Integrin beta 1 is crucial for urinary concentrating ability and renal medulla architecture in adult mice
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Anna Iervolino, Luigi R. De La Motte, Federica Petrillo, Federica Prosperi, Francesca Maria Alvino, Guglielmo Schiano, Alessandra F. Perna, Danilo Di Matteo, Mario De Felice, Giovambattista Capasso, Francesco Trepiccione, Iervolino, Anna, De La Motte, Luigi R, Petrillo, Federica, Prosperi, Federica, Schiano, Guglielmo, Perna, Alessandra F, Di Matteo, Danilo, De Felice, Mario, Capasso, Giovambattista, Trepiccione, Francesco, Iervolino, A., De La Motte, L. R., Petrillo, F., Prosperi, F., Schiano, G., Perna, A. F., Di Matteo, D., De Felice, M., Capasso, G., and Trepiccione, F.
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0301 basic medicine ,Collecting duct ,medicine.medical_specialty ,Renal failure ,Physiology ,Urinary system ,Integrin ,030232 urology & nephrology ,AQP2 ,collecting duct ,integrin beta1 ,renal failure ,thick ascending limb ,urologic and male genital diseases ,lcsh:Physiology ,Extracellular matrix ,Beta-1 adrenergic receptor ,03 medical and health sciences ,0302 clinical medicine ,Cell–cell interaction ,Polyuria ,Physiology (medical) ,Internal medicine ,Renal medulla ,medicine ,Anoikis ,Thick ascending limb ,lcsh:QP1-981 ,biology ,business.industry ,urogenital system ,Integrin beta1 ,Correction ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,Aquaporin 2 ,biology.protein ,medicine.symptom ,business ,030217 neurology & neurosurgery - Abstract
Integrins are heterodimers anchoring cells to the surrounding extracellular matrix (ECM), an active and complex process mediating a series of inside-out and outside-in stimuli regulating cellular turn-over, tissue growth and architecture. Itgb1 is the main subunit of the renal integrins and it is critical for renal development. This study aims to investigate the role of Itgb1 in the adult renal epithelial cells by knocking down Itgb1 in PAX8 expressing cells. Itgb1-Pax8 cKO mice develop a progressively worsening proteinuria and renal abnormalities leading to severe renal failure and hypertension. This phenotype is also associated with severe dysfunction of distal nephron and polyuria. To further investigate whether distal nephron involvement was primarily related to Itgb1 suppression or secondary to renal failure, an Itgb1-AQP2 cKO mouse model was generated. These mice lack Itgb1 expression in AQP2 expressing cells. They do not show any developmental alteration, but 1 month old mice are resistant to dDAVP administration and finally, at 2 months of age, they develop overt polyuria. This phenotype is due to primary collecting duct (CD) cells anoikis. The entire architecture of the outer medulla is altered, with loss of the typical organization pattern of vascular and tubular bundles alternation. Indeed, even though not primarily affected by genetic ablation, the TAL is secondarily affected in this model. It is sufficient to suppress Itgb1 expression in the CD in order to stimulate proliferation and then disappearance of neighboring TAL cells. This study shows that cell to cell interaction through the ECM is critical for architecture and function maintenance of the outer medulla and that Itgb1 is crucial for this process. Integrins are heterodimers anchoring cells to the surrounding extracellular matrix (ECM), an active and complex process mediating a series of inside-out and outside-in stimuli regulating cellular turn-over, tissue growth and architecture. Itgb1 is the main subunit of the renal integrins and it is critical for renal development. This study aims to investigate the role of Itgb1 in the adult renal epithelial cells by knocking down Itgb1 in PAX8 expressing cells. Itgb1-Pax8 cKO mice develop a progressively worsening proteinuria and renal abnormalities leading to severe renal failure and hypertension. This phenotype is also associated with severe dysfunction of distal nephron and polyuria. To further investigate whether distal nephron involvement was primarily related to Itgb1 suppression or secondary to renal failure, an Itgb1-AQP2 cKO mouse model was generated. These mice lack Itgb1 expression in AQP2 expressing cells. They do not show any developmental alteration, but 1 month old mice are resistant to dDAVP administration and finally, at 2 months of age, they develop overt polyuria. This phenotype is due to primary collecting duct (CD) cells anoikis. The entire architecture of the outer medulla is altered, with loss of the typical organization pattern of vascular and tubular bundles alternation. Indeed, even though not primarily affected by genetic ablation, the TAL is secondarily affected in this model. It is sufficient to suppress Itgb1 expression in the CD in order to stimulate proliferation and then disappearance of neighboring TAL cells. This study shows that cell to cell interaction through the ECM is critical for architecture and function maintenance of the outer medulla and that Itgb1 is crucial for this process.
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- 2018
15. 'Stockpile' of Slight Transcriptomic Changes Determines the Indirect Genotoxicity of Low-Dose BPA in Thyroid Cells
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Concetta Ambrosino, Danila Cuomo, Immacolata Porreca, Massimo Mallardo, Mario De Felice, Luisa Ulloa Severino, Michele Ceccarelli, Angela Nebbioso, Elena Amendola, Lucia Altucci, Fulvio D'Angelo, Porreca, I, Ulloa Severino, L, D'Angelo, F, Cuomo, D, Ceccarelli, M, Altucci, L, Amendola, E, Nebbioso, A, Mallardo, M, De Felice, M, Ambrosino, C., Porreca, Immacolata, ULLOA SEVERINO, Luisa, D'Angelo, Fulvio, Cuomo, Danila, Ceccarelli, Michele, Altucci, Lucia, Amendola, Elena, Nebbioso, Angela, Mallardo, Massimo, De Felice, Mario, Ambrosino, Concetta, and Ulloa Severino, Luisa
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0301 basic medicine ,Genetics and Molecular Biology (all) ,Time Factors ,Light ,Benzhydryl Compounds ,Cell Line ,DNA Damage ,DNA Replication ,Dose-Response Relationship, Drug ,Gene Expression ,Humans ,Phenols ,Thyroid Gland ,Transcriptome ,Medicine (all) ,Biochemistry, Genetics and Molecular Biology (all) ,Agricultural and Biological Sciences (all) ,Microarrays ,DNA rapair ,lcsh:Medicine ,010501 environmental sciences ,medicine.disease_cause ,01 natural sciences ,Biochemistry ,Medicine and Health Sciences ,lcsh:Science ,Benzhydryl Compound ,Thyroid ,Multidisciplinary ,Physics ,Genomics ,Cell biology ,Nucleic acids ,Chemistry ,Bioassays and Physiological Analysis ,Toxicity ,Physical Sciences ,DNA microarray ,Drug ,Anatomy ,Transcriptome Analysis ,hormones, hormone substitutes, and hormone antagonists ,Human ,Research Article ,Ultraviolet radiation ,Programmed cell death ,endocrine system ,Pollutants ,Time Factor ,DNA repair ,DNA damage ,DNA recombination ,Endocrine System ,Biology ,Research and Analysis Methods ,Dose-Response Relationship ,03 medical and health sciences ,Electromagnetic radiation ,medicine ,Genetics ,Environmental Chemistry ,0105 earth and related environmental sciences ,Phenol ,Biology and life sciences ,urogenital system ,lcsh:R ,Ecology and Environmental Sciences ,Computational Biology ,DNA Repair Pathway ,DNA ,Genome Analysis ,Molecular biology ,030104 developmental biology ,lcsh:Q ,Ultraviolet C ,Genotoxicity - Abstract
Epidemiological and experimental data highlighted the thyroid-disrupting activity of bisphenol A (BPA). Although pivotal to identify the mechanisms of toxicity, direct low-dose BPA effects on thyrocytes have not been assessed. Here, we report the results of microarray experiments revealing that the transcriptome reacts dynamically to low-dose BPA exposure, adapting the changes in gene expression to the exposure duration. The response involves many genes, enriching specific pathways and biological functions mainly cell death/proliferation or DNA repair. Their expression is only slightly altered but, since they enrich specific pathways, this results in major effects as shown here for transcripts involved in the DNA repair pathway. Indeed, even though no phenotypic changes are induced by the treatment, we show that the exposure to BPA impairs the cell response to further stressors. We experimentally verify that prolonged exposure to low doses of BPA results in a delayed response to UV-C-induced DNA damage, due to impairment of p21-Tp53 axis, with the BPA-treated cells more prone to cell death and DNA damage accumulation. The present findings shed light on a possible mechanism by which BPA, not able to directly cause genetic damage at environmental dose, may exert an indirect genotoxic activity.
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- 2016
16. A novel DNA helicase with strand-annealing activity from the crenarchaeon Sulfolobus solfataricus
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Francesca M. Pisani, Mariarita De Felice, Valentina Aria, Mariarosaria De Falco, Biagio Pucci, Luca Esposito, Mosè Rossi, De Felice, M., Aria, V., Esposito, L., De Falco, M., Pucci, B., Rossi, Mose', and Pisani, F. M.
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DNA recombination ,ved/biology.organism_classification_rank.species ,DNA helicase ,Biochemistry ,Catalysis ,Substrate Specificity ,law.invention ,chemistry.chemical_compound ,Adenosine Triphosphate ,law ,Annealing activity ,Molecular Biology ,biology ,ved/biology ,Hydrolysis ,Circular bacterial chromosome ,Sulfolobus solfataricus ,DNA Helicases ,DNA replication ,Helicase ,DNA ,Cell Biology ,Archaea ,DNA-pairing activity ,chemistry ,biology.protein ,Recombinant DNA ,Primase ,Dimerization ,genome stability ,Research Article ,Protein Binding - Abstract
To protect their genetic material cells adopt different mechanisms linked to DNA replication, recombination and repair. Several proteins function at the interface of these DNA transactions. In the present study, we report on the identification of a novel archaeal DNA helicase. BlastP searches of the Sulfolobus solfataricus genome database allowed us to identify an open reading frame (SSO0112, 875 amino acid residues) having sequence similarity with the human RecQ5β. The corresponding protein, termed Hel112 by us, was produced in Escherichia coli in soluble form, purified to homogeneity and characterized. Gel-filtration chromatography and glycerol-gradient sedimentation analyses revealed that Hel112 forms monomers and dimers in solution. Biochemical characterization of the two oligomeric species revealed that only the monomeric form has an ATP-dependent 3′–5′ DNA-helicase activity, whereas, unexpectedly, both the monomeric and dimeric forms possess DNA strand-annealing capability. The Hel112 monomeric form is able to unwind forked and 3′-tailed DNA structures with high efficiency, whereas it is almost inactive on blunt-ended duplexes and bubble-containing molecules. This analysis reveals that S. solfataricus Hel112 shares some enzymatic features with the RecQ-like DNA helicases and suggests potential cellular functions of this protein.
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- 2007
17. Evaluation of low doses BPA-induced perturbation of glycemia by toxicogenomics points to a primary role of pancreatic islets and to the mechanism of toxicity
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Immacolata Porreca, Concetta Ambrosino, Fulvio D'Angelo, M De Felice, Danila Cuomo, Massimo Mallardo, Emanuele Carchia, P. J. Almeida, Michele Ceccarelli, Carchia, E, Porreca, I, Almeida, Pj, D'Angelo, F, Cuomo, D, Ceccarelli, M, De Felice, M, Mallardo, M, and Ambrosino, C.
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Blood Glucose ,Male ,Cancer Research ,medicine.medical_specialty ,endocrine system ,Immunology ,BPA-induced ,glycemia ,toxicogenomics ,pancreatic ,toxicity ,Biology ,Toxicogenetics ,Islets of Langerhans ,Mice ,Cellular and Molecular Neuroscience ,Phenols ,In vivo ,Internal medicine ,medicine ,Animals ,Homeostasis ,Glucose homeostasis ,Benzhydryl Compounds ,geography ,geography.geographical_feature_category ,Dose-Response Relationship, Drug ,Pancreatic islets ,Cell Biology ,Islet ,Mice, Inbred C57BL ,Endocrinology ,medicine.anatomical_structure ,Toxicity ,Hepatocytes ,Original Article ,Toxicogenomics ,Ex vivo ,hormones, hormone substitutes, and hormone antagonists - Abstract
Epidemiologic and experimental studies have associated changes of blood glucose homeostasis to Bisphenol A (BPA) exposure. We took a toxicogenomic approach to investigate the mechanisms of low-dose (1 × 10−9M) BPA toxicity in ex vivo cultures of primary murine pancreatic islets and hepatocytes. Twenty-nine inhibited genes were identified in islets and none in exposed hepatocytes. Although their expression was slightly altered, their impaired cellular level, as a whole, resulted in specific phenotypic changes. Damage of mitochondrial function and metabolism, as predicted by bioinformatics analyses, was observed: BPA exposure led to a time-dependent decrease in mitochondrial membrane potential, to an increase of ROS cellular levels and, finally, to an induction of apoptosis, attributable to the bigger Bax/Bcl-2 ratio owing to activation of NF-κB pathway. Our data suggest a multifactorial mechanism for BPA toxicity in pancreatic islets with emphasis to mitochondria dysfunction and NF-κB activation. Finally, we assessed in vitro the viability of BPA-treated islets in stressing condition, as exposure to high glucose, evidencing a reduced ability of the exposed islets to respond to further damages. The result was confirmed in vivo evaluating the reduction of glycemia in hyperglycemic mice transplanted with control and BPA-treated pancreatic islets. The reported findings identify the pancreatic islet as the main target of BPA toxicity in impairing the glycemia. They suggest that the BPA exposure can weaken the response of the pancreatic islets to damages. The last observation could represent a broader concept whose consideration should lead to the development of experimental plans better reproducing the multiple exposure conditions.
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- 2015
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18. Selective Dicer Suppression in the Kidney Alters gsk3?/?-Catenin Pathways Promoting a Glomerulocystic Disease
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Manuela Spagnuolo, Mario De Felice, Federica Petrillo, Marzia Scarfò, Gabriella De Vita, Daniela Frezzetti, Francesco Trepiccione, Giovambattista Capasso, Anna Iervolino, Iervolino, A, Trepiccione, Francesco, Petrillo, F, Spagnuolo, M, Scarfò, M, Frezzetti, D, De Vita, G, De Felice, M, Capasso, Giovambattista, Iervolino, Anna, Petrillo, Federica, Spagnuolo, Manuela, Scarfò, Marzia, Frezzetti, Daniela, DE VITA, Gabriella, and DE FELICE, Mario
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Ribonuclease III ,medicine.medical_specialty ,Renal cortex ,lcsh:Medicine ,Biology ,Kidney ,DEAD-box RNA Helicases ,Glycogen Synthase Kinase 3 ,Mice ,PAX8 Transcription Factor ,Downregulation and upregulation ,Internal medicine ,Conditional gene knockout ,medicine ,Animals ,Paired Box Transcription Factors ,lcsh:Science ,beta Catenin ,Mice, Knockout ,Glycogen Synthase Kinase 3 beta ,Multidisciplinary ,Cilium ,lcsh:R ,Kidney metabolism ,Kidney Diseases, Cystic ,Phenotype ,Endocrinology ,medicine.anatomical_structure ,Gene Expression Regulation ,biology.protein ,Cancer research ,lcsh:Q ,Research Article ,Signal Transduction ,Dicer - Abstract
Dicer is a crucial enzyme for the maturation of miRNAs. Mutations in the Dicer gene are highly associated with Pleuro Pulmonary Blastoma-Family Dysplasia Syndrome (PPB-FDS, OMIM 601200), recently proposed to be renamed Dicer syndrome. Aside from the pulmonary phenotype (blastoma), renal nephroma and thyroid goiter are frequently part of Dicer syndrome. To investigate the renal phenotype, conditional knockout (cKO) mice for Dicer in Pax8 expressing cells were generated. Dicer cKO mice progressively develop a glomerulocystic phenotype coupled with urinary concentration impairment, proteinuria and severe renal failure. Higher cellular turnover of the parietal cells of Bowman's capsule precedes the development of the cysts and the primary cilium progressively disappears with cyst-enlargement. Upregulation of GSK3β precedes the development of the glomerulocystic phenotype. Downregulation of β-catenin in the renal cortex and its cytosolic removal in the cells lining the cysts may be associated with observed accumulation of GSK3β. Alterations of β-catenin regulating pathways could promote cystic degeneration as in other models. Thus, miRNAs are fundamental in preserving renal morphology and function. Alteration of the GSK3β/β-catenin pathway could be a crucial mechanism linking miRNA dysregulation and the development of a glomerulocystic disease.
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- 2015
19. The human GINS complex binds to and specifically stimulates human DNA polymerase α‐primase
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Mariarita De Felice, Mariarosaria De Falco, Francesca M. Pisani, Ulrich Hübscher, Mosè Rossi, Elena Ferrari, De Falco, M, Ferrari, E, De Felice, M, Rossi, Mose', Hubscher, U, and Pisani, F. M.
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GINS ,DNA polymerase ,Scientific Report ,DNA replication ,Biochemistry ,genome dynamics ,law.invention ,chemistry.chemical_compound ,ATP Binding Cassette Transporter, Subfamily B, Member 3 ,law ,Escherichia coli ,Genetics ,Humans ,ATP Binding Cassette Transporter, Subfamily B, Member 2 ,Protein Precursors ,Molecular Biology ,Polymerase ,DNA primase ,biology ,DNA ,Surface Plasmon Resonance ,DNA Polymerase I ,Molecular biology ,Peptide Fragments ,Recombinant Proteins ,Cell biology ,Protein Subunits ,chemistry ,Recombinant DNA ,biology.protein ,ATP-Binding Cassette Transporters ,Primase ,DNA polymerase I - Abstract
The eukaryotic GINS complex has an essential role in the initiation and elongation phases of genome duplication. It is composed of four paralogous subunits--Sld5, Psf1, Psf2 and Psf3--which are ubiquitous and evolutionarily conserved in eukaryotic organisms. Here, we report the biochemical characterization of the human GINS complex (hGINS). The four hGINS subunits were coexpressed in Escherichia coli in a highly soluble form and purified as a complex. hGINS was shown to interact directly with the heterodimeric human DNA primase, by using either surface plasmon resonance measurements or by immunoprecipitation experiments carried out with anti-hGINS antibodies. The DNA polymerase alpha-primase synthetic activity was specifically stimulated by hGINS on various primed DNA templates. The significance of these findings is discussed in view of the molecular dynamics at the human replication fork.
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- 2006
20. Alteration of cell morphology and viability in a recA mutant of Streptococcus thermophilus upon induction of heat shock and nutrient starvation
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Ezio Ricca, Gino Naclerio, Gabriele Giliberti, Maurilio De Felice, Luca Martirani, Giliberti, G., Naclerio, G., Martirani, L, Ricca, Ezio, DE FELICE, M., Giliberti, G, Naclerio, G, Ricca, E, and DE FELICE, Maurilio
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Streptococcus thermophilus ,Hot Temperature ,Time Factors ,Mutant ,Cell morphology ,Bacterial Proteins ,Heat shock protein ,Gene expression ,Genetics ,Anaerobiosis ,biology ,Streptococcus ,Chaperonin 60 ,General Medicine ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,GroEL ,Erythromycin ,Rec A Recombinases ,Biochemistry ,Mutation ,Microscopy, Electron, Scanning ,bacteria ,Electrophoresis, Polyacrylamide Gel ,Cell Division ,Bacteria ,Intracellular ,Molecular Chaperones - Abstract
We identified the recA gene of the moderately thermophilic bacterium Streptococcus thermophilus and investigated the role of its product in the adaptation to heat shock and nutrient starvation. Expression of recA was required for optimal viability and normal cell morphology upon induction of both stresses. Normal induction of GroEL and ClpL in a recA knock-out mutant suggests that the RecA role in heat shock and nutrient starvation response of S. thermophilus is independent from the intracellular accumulation of these stress-specific chaperones.
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- 2002
21. Comparative genomics reveals a functional thyroid-specific element in the far upstream region of the PAX8 gene
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Tiziana de Cristofaro, Mariastella Zannini, Roberto Nitsch, Roberto Di Lauro, Alessandra di Gennaro, Mario De Felice, Serena Abbondante, Valeria Di Dato, Nitsch, R., Di Dato, V., di Gennaro, A., de Cristofaro, T., Abbondante, S., De Felice, M., Zannini, M., and Di Lauro, R.
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Transcription, Genetic ,endocrine system diseases ,PROMOTER ,5' Flanking Region ,medicine.medical_treatment ,Thyroid Gland ,Mice ,0302 clinical medicine ,Paired Box Transcription Factors ,TRANSCRIPTION FACTOR ,Conserved Sequence ,Phylogeny ,Genetics ,Regulation of gene expression ,0303 health sciences ,biology ,HOMEODOMAIN ,Genomics ,DNA-Binding Proteins ,DIFFERENTIATION ,Enhancer Elements, Genetic ,Organ Specificity ,030220 oncology & carcinogenesis ,RECEPTOR ACTIVATOR ,Research Article ,Biotechnology ,endocrine system ,lcsh:QH426-470 ,lcsh:Biotechnology ,5' flanking region ,BAC DNA ,ENHANCER ,PAX8 Transcription Factor ,03 medical and health sciences ,Thyroid peroxidase ,lcsh:TP248.13-248.65 ,Sequence Homology, Nucleic Acid ,medicine ,Animals ,Humans ,Enhancer ,Transcription factor ,030304 developmental biology ,RECOGNITION ,lcsh:Genetics ,Gene Expression Regulation ,CELLS ,biology.protein ,Thyroglobulin ,PAX8 ,Sequence Alignment ,TISSUE-SPECIFIC EXPRESSION ,Transcription Factors ,FOXE1 - Abstract
Background The molecular mechanisms leading to a fully differentiated thyrocite are still object of intense study even if it is well known that thyroglobulin, thyroperoxidase, NIS and TSHr are the marker genes of thyroid differentiation. It is also well known that Pax8, TTF-1, Foxe1 and Hhex are the thyroid-enriched transcription factors responsible for the expression of the above genes, thus are responsible for the differentiated thyroid phenotype. In particular, the role of Pax8 in the fully developed thyroid gland was studied in depth and it was established that it plays a key role in thyroid development and differentiation. However, to date the bases for the thyroid-enriched expression of this transcription factor have not been unraveled yet. Here, we report the identification and characterization of a functional thyroid-specific enhancer element located far upstream of the Pax8 gene. Results We hypothesized that regulatory cis-acting elements are conserved among mammalian genes. Comparison of a genomic region extending for about 100 kb at the 5'-flanking region of the mouse and human Pax8 gene revealed several conserved regions that were tested for enhancer activity in thyroid and non-thyroid cells. Using this approach we identified one putative thyroid-specific regulatory element located 84.6 kb upstream of the Pax8 transcription start site. The in silico data were verified by promoter-reporter assays in thyroid and non-thyroid cells. Interestingly, the identified far upstream element manifested a very high transcriptional activity in the thyroid cell line PC Cl3, but showed no activity in HeLa cells. In addition, the data here reported indicate that the thyroid-enriched transcription factor TTF-1 is able to bind in vitro and in vivo the Pax8 far upstream element, and is capable to activate transcription from it. Conclusions Results of this study reveal the presence of a thyroid-specific regulatory element in the 5' upstream region of the Pax8 gene. The identification of this regulatory element represents the first step in the investigation of upstream regulatory mechanisms that control Pax8 transcription during thyroid differentiation and are relevant to further studies on Pax8 as a candidate gene for thyroid dysgenesis.
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- 2010
22. Isolation of an Escherichia coli K4 kfoC mutant over-producing capsular chondroitin
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Mario De Rosa, Eugenio Notomista, Donatella Cimini, Chiara Schiraldi, Mario Varcamonti, Odile Francesca Restaino, Maurilio De Felice, Anna Zanfardino, Zanfardino, A., Restaino, Odile Francesca, Notomista, E., Cimini, Donatella, Schiraldi, Chiara, DE ROSA, Mario, De Felice, M., Varcamonti, M., Zanfardino, Anna, Restaino, Of, Notomista, Eugenio, Cimini, D, Schiraldi, C, De Rosa, M, DE FELICE, Maurilio, and Varcamonti, Mario
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Bacterial capsule ,Mutant ,Mutation, Missense ,lcsh:QR1-502 ,Mutagenesis (molecular biology technique) ,Bioengineering ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,lcsh:Microbiology ,Microbiology ,chemistry.chemical_compound ,medicine ,Escherichia coli ,Chondroitin ,Bacterial Capsules ,chemistry.chemical_classification ,biology ,Organisms, Genetically Modified ,Escherichia coli Proteins ,Research ,Anti-Inflammatory Agents, Non-Steroidal ,Chondroitin Sulfates ,Wild type ,Gene Expression Regulation, Bacterial ,biology.organism_classification ,carbohydrates (lipids) ,Enzyme ,chemistry ,Biochemistry ,Hexosyltransferases ,Bacteria ,Biotechnology - Abstract
Background Chondroitin sulphate is a complex polysaccharide having important structural and protective functions in animal tissues. Extracted from animals, this compound is used as a human anti-inflammatory drug. Among bacteria, Escherichia coli K4 produces a capsule containing a non-sulphate chondroitin and its development may provide an efficient and cheap fermentative production of the polysaccharide. Results A random N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis was performed on E. coli K4 to isolate mutants showing an increased production of chondroitin. Several mutants were isolated, one of which, here named VZ15, produced about 80% more chondroitin than the wild type E. coli. We found that the mutant has a missense mutation in the codon 313 of kfoC, the gene encoding chondroitin polymerase (K4CP), with a change from arginine to glutamine. A docking analysis to explain the increased productivity of the K4CP enzyme is presented. Conclusion The enhanced chondroitin production by the E. coli K4 mutant reported here shows the validity of the strain improvement strategy for more cost-friendly fermentative processes in the production of this pharmaceutically important but so-far expensive polysaccharide.
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- 2010
23. Conditional inactivation of the E-cadherin gene in thyroid follicular cells affects gland development but does not impair junction formation
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Barbara D'Andrea, Günther Schütz, Lucio Nitsch, Tim M. Wintermantel, Oreda Boussadia, Roberto Di Lauro, Gaetano Calì, Rolf Kemler, Mariastella Zannini, Daniela Terracciano, Mario De Felice, Andrea Affuso, Daniel Silberschmidt, Elena Amendola, Patrizia Rubini, Carlo Tacchetti, Cali', G., Zannini, M., Rubini, P., Tacchetti, Carlo, Dandrea, B., Affuso, A., Wintermantel, T., Boussadia, O., Terracciano, D., Silberschmidt, D. ., Amendola, E, DE FELICE, M., Schtz, G., Kemler, R., DI LAURO, R., Nitsch, L., Calì, G., Tacchetti, C., D'Andrea, Barbara, Terracciano, Daniela, Silberschmidt, D., Amendola, Elena, DE FELICE, Mario, Schütz, G., DI LAURO, Roberto, and Nitsch, Lucio
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Male ,medicine.medical_specialty ,endocrine system ,medicine.medical_treatment ,Thyroid Gland ,Biology ,Follicular cell ,Tight Junctions ,thyroid ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Pregnancy ,Occludin ,Internal medicine ,Claudin-1 ,Conditional gene knockout ,Cell Adhesion ,medicine ,Animals ,junction ,beta Catenin ,030304 developmental biology ,Mice, Knockout ,0303 health sciences ,Tight junction ,Cadherin ,Gene Expression Profiling ,Thyroid ,Gene Expression Regulation, Developmental ,Membrane Proteins ,E-cadherin ,Cadherins ,Phosphoproteins ,Mice, Inbred C57BL ,adhesion ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Knockout mouse ,Zonula Occludens-1 Protein ,Female ,Thyroglobulin ,Endocrine gland - Abstract
We have conditionally inactivated the E-cadherin gene in the thyroid follicular cells of mouse embryo to unravel its role in thyroid development. We used the Cre-loxP system in which the Cre-recombinase was expressed under the control of the tissue-specific thyroglobulin promoter that becomes active at embryonic d 15. At postnatal d 7, thyroid follicle lumens in the knockout mice were about 30% smaller with respect to control mice and had an irregular shape. E-cadherin was almost completely absent in thyrocytes, beta-catenin was significantly reduced, whereas no change in gamma-catenin was detected. alpha-Catenin was also reduced on the cell plasma membrane. Despite the dramatic loss of E-cadherin and beta-catenin, cell-cell junctions were not affected, the distribution of tight junction proteins was unaltered, and no increase of thyroglobulin circulating in the blood was observed. In addition, we found that other members of the cadherin family, the R-cadherin and the Ksp-cadherin, were expressed in thyrocytes and that their membrane distribution was not altered in the E-cadherin conditional knockout mouse. Our results indicate that E-cadherin has a role in the development of the thyroid gland and in the expression of beta-catenin, but it is not essential for the maintenance of follicular cell adhesion.
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- 2007
24. Modular organization of the Sulfolobus solfataricus mini-chromosome maintenance protein
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Monia Rocco, Mariarita De Felice, Francesco Esposito, Mariarosaria De Falco, Mosè Rossi, Francesca M. Pisani, Luca Esposito, Biagio Pucci, Pucci, B, DE FELICE, M, Rocco, M, Esposito, F, DE FALCO, M, Esposito, L, Rossi, Mose', and Pisani, Fm
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Archaeal Proteins ,ved/biology.organism_classification_rank.species ,Biochemistry ,Models, Biological ,chemistry.chemical_compound ,Structure-Activity Relationship ,Protein structure ,Minichromosome maintenance ,MCM complex ,Amino Acid Sequence ,Molecular Biology ,Sequence Deletion ,Adenosine Triphosphatases ,biology ,ved/biology ,Oligonucleotide ,Sulfolobus solfataricus ,DNA Helicases ,Helicase ,Cell Biology ,DNA Replication Fork ,Recombinant Proteins ,Protein Structure, Tertiary ,DNA-Binding Proteins ,chemistry ,biology.protein ,DNA - Abstract
Mini-chromosome maintenance (MCM) proteins form ring-like hexameric complexes that are commonly believed to act as the replicative DNA helicase at the eukaryotic/archaeal DNA replication fork. Because of their simplified composition with respect to the eukaryotic counterparts, the archaeal MCM complexes represent a good model system to use in analyzing the structural/functional relationships of these important replication factors. In this study the domain organization of the MCM-like protein from Sulfolobus solfataricus (Sso MCM) has been dissected by trypsin partial proteolysis. Three truncated derivatives of Sso MCM corresponding to protease-resistant domains were produced as soluble recombinant proteins and purified: the N-terminal domain (N-ter, residues 1-268); a fragment comprising the AAA+ and C-terminal domains (AAA+-C-ter, residues 269-686); and the C-terminal domain (C-ter, residues 504-686). All of the purified recombinant proteins behaved as monomers in solution as determined by analytical gel filtration chromatography, suggesting that the polypeptide chain integrity is required for stable oligomerization of Sso MCM. However, the AAA+-C-ter derivative, which includes the AAA+ motor domain and retains ATPase activity, was able to form dimers in solution when ATP was present, as analyzed by size exclusion chromatography and glycerol gradient sedimentation analyses. Interestingly, the AAA+-C-ter protein could displace oligonucleotides annealed to M13 single-stranded DNA although with a reduced efficiency in comparison with the full-sized Sso MCM. The implications of these findings for understanding the DNA helicase mechanism of the MCM complex are discussed.
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- 2007
25. Biochemical evidence of a physical interaction between Sulfolobus solfataricus B-family and Y-family DNA polymerases
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Francesca M. Pisani, Biagio Pucci, Mariarosaria De Falco, Mariarita De Felice, Petr Grúz, Luca Esposito, Takehiko Nohmi, Mosè Rossi, Barbara Medagli, De Felice, M., Medagli, B., Esposito, L., De Falco, M., Pucci, B., Rossi, Mose', Gruz, P., Nohmi, T., and Pisani, F. M.
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DNA polymerase ,Archaeal Proteins ,DNA polymerase II ,ved/biology.organism_classification_rank.species ,DNA-Directed DNA Polymerase ,DNA replication ,Microbiology ,DNA polymerase delta ,Protein Structure, Secondary ,Genome, Archaeal ,Genome stability ,Translesion synthesis ,biology ,ved/biology ,Sulfolobus solfataricus ,General Medicine ,Base excision repair ,Processivity ,Surface Plasmon Resonance ,Archaea ,Biochemistry ,Coding strand ,biology.protein ,Molecular Medicine ,DNA Damage ,Protein Binding - Abstract
The hyper-thermophilic archaeon Sulfolobus solfataricus possesses two functional DNA polymerases belonging to the B-family (Sso DNA pol B1) and to the Y-family (Sso DNA pol Y1). Sso DNA pol B1 recognizes the presence of uracil and hypoxanthine in the template strand and stalls synthesis 3-4 bases upstream of this lesion ("read-ahead" function). On the other hand, Sso DNA pol Y1 is able to synthesize across these and other lesions on the template strand. Herein we report evidence that Sso DNA pol B1 physically interacts with DNA pol Y1 by surface plasmon resonance measurements and immuno-precipitation experiments. The region of DNA pol B1 responsible for this interaction has been mapped in the central portion of the polypeptide chain (from the amino acid residue 482 to 617), which includes an extended protease hyper-sensitive linker between the N- and C-terminal modules (amino acid residues Asn482-Ala497) and the alpha-helices forming the "fingers" sub-domain (alpha-helices R, R' and S). These results have important implications for understanding the polymerase-switching mechanism on the damaged template strand during genome replication in S. solfataricus.
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- 2007
26. Replacement of K-Ras with H-Ras supports normal embryonic development despite inducing cardiovascular pathology in adult mice
- Author
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Carmine Vecchione, Antonella Notte, Tommaso Russo, Nicoletta Potenza, Annamaria Rosica, Giuseppe Lembo, Mario De Felice, Roberto Di Lauro, Assunta De Rienzo, Andrea Affuso, Giuseppe Cifelli, Gabriella De Vita, Roberta Poulet, Lisa Bauer, Potenza, Nicoletta, Vecchione, C, Notte, A, DE RIENZO, A, Rosica, A, Bauer, L, Affuso, A, DE FELICE, M, Russo, T, Poulet, R, Cifelli, G, DE VITA, G, Lembo, G, DI LAURO, R., Potenza, N., Vecchione, C., Notte, A., DE RIENZO, A., Rosica, A., Bauer, L., Affuso, A., DE FELICE, Mario, Russo, Tommaso, Poulet, R., Cifelli, G., DE VITA, Gabriella, Lembo, G., and DI LAURO, Roberto
- Subjects
Cardiomyopathy, Dilated ,medicine.medical_specialty ,pressione arteriosa ,Cardiomyopathy ,Scientific Report ,GTPase ,Biology ,medicine.disease_cause ,Biochemistry ,cardiomiopatia ,K-ra ,symbols.namesake ,Mice ,Internal medicine ,Genetics ,medicine ,Animals ,Molecular Biology ,Gene ,ras ,Embryogenesis ,h-ras ,medicine.disease ,Phenotype ,Mice, Mutant Strains ,H-ra ,Cell biology ,k-ras ,Endocrinology ,Genes, ras ,Hypertension ,Embryonic development ,Mendelian inheritance ,symbols ,Blood pressure ,Carcinogenesis ,Function (biology) ,blood pressure ,cardiomyopathy ,embryonic development - Abstract
Ras proteins are highly related GTPases that have key roles in regulating growth, differentiation and tumorigenesis. Gene-targeting experiments have shown that, out of the three mammalian ras genes, only K-ras is essential for normal mouse embryogenesis, and that mice deprived of H-ras and/or N-ras show no major phenotype. We generated mice (HrasKI) in which the K-ras gene had been modified to encode H-Ras protein. HrasKI mice produce undetectable amounts of K-Ras but - in contrast to mice homozygous for a null K-ras allele - they are born at the expected mendelian frequency, indicating that H-Ras can be substituted for K-Ras in embryonic development. However, adult HrasKI mice show dilated cardiomyopathy associated with arterial hypertension. Our results show that K-Ras can be replaced by H-Ras in its essential function in embryogenesis, and indicate that K-Ras has a unique role in cardiovascular homeostasis. © 2005 European Molecular Biology Organization.
- Published
- 2005
27. GerE-independent expression of cotH leads to CotC accumulation in the mother cell compartment during Bacillus subtilis sporulation
- Author
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Gaetano Castaldo, Giuseppina Cangiano, Ezio Ricca, Loredana Baccigalupi, Maurilio De Felice, Rosangela Marasco, Rachele Isticato, Baccigalupi, L., Castaldo, G., Cangiano, G., Istigato, R., Marasco, Rosangela, DE FELICE, M., Ricca, E., Baccigalupi, Loredana, Castaldo, G, Cangiano, Giuseppina, Isticato, Rachele, Marasco, R, DE FELICE, Maurilio, and Ricca, Ezio
- Subjects
Spores, Bacterial ,Mutation ,sporulation ,Bacillaceae ,biology ,fungi ,cotH ,Promoter ,Bacillus subtilis ,Gene Expression Regulation, Bacterial ,biology.organism_classification ,medicine.disease_cause ,Microbiology ,Cell biology ,Gene product ,Bacterial Proteins ,Gene expression ,Transcriptional regulation ,medicine ,Bacillus subtili ,Gene - Abstract
The cotG and cotH genes of Bacillus subtilis encode two previously characterized spore coat proteins. The two genes are adjacent on the chromosome and divergently transcribed by sigma(K), a sporulation-specific sigma factor of the RNA polymerase. We report evidence that the cotH promoter maps 812 bp upstream of the beginning of its coding region and that the divergent cotG gene is entirely contained between the promoter and the coding part of cotH. A bioinformatic analysis of all entirely sequenced prokaryotic genomes showed that such chromosomal organization is not common in spore-forming bacilli. Indeed, CotG is present only in B. subtilis, B. amyloliquefaciens, and B. atrophaeus and in two Geobacillus strains. When present, cotG always encodes a modular protein composed of tandem repeats and is always close to but divergently transcribed with respect to cotH. Bioinformatic and phylogenic data suggest that such genomic organizations have a common evolutionary origin and that the modular structure of the extant cotG genes is the outcome of multiple rounds of gene elongation events of an ancestral minigene
- Published
- 2004
28. Biochemical characterization of a CDC6-like protein from the crenarchaeon Sulfolobus solfataricus
- Author
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Floriana Carpentieri, Francesca M. Pisani, Luca Esposito, Mariarita De Felice, Biagio Pucci, Mariarosaria De Falco, Mosè Rossi, DE FELICE, M, Esposito, L, Pucci, B, Carpentieri, F, DE FALCO, M, Rossi, Mose', and Pisani, Fm
- Subjects
Saccharomyces cerevisiae Proteins ,Archaeal Proteins ,ved/biology.organism_classification_rank.species ,Mutation, Missense ,Sequence Homology ,Cell Cycle Proteins ,Biology ,Cdc6 ,Biochemistry ,Sulfolobus ,chemistry.chemical_compound ,Adenosine Triphosphate ,MCM complex ,Cloning, Molecular ,Phosphorylation ,Molecular Biology ,Gene ,Replicazione del DNA ,ved/biology ,Autophosphorylation ,Sulfolobus solfataricus ,Walker motifs ,MCM ,Cell Biology ,Archaea ,MCM Protein ,DNA-Binding Proteins ,DNA helicase activity ,chemistry ,DNA - Abstract
Cdc6 proteins play an essential role in the initiation of chromosomal DNA replication in Eukarya. Genes coding for putative homologs of Cdc6 have been also identified in the genomic sequence of Archaea, but the properties of the corresponding proteins have been poorly investigated so far. Herein, we report the biochemical characterization of one of the three putative Cdc6-like factors from the hyperthermophilic crenarchaeon Sulfolobus solfataricus (SsoCdc6-1). SsoCdc6-1 was overproduced in Escherichia coli as a His-tagged protein and purified to homogeneity. Gel filtration and glycerol gradient ultracentrifugation experiments indicated that this protein behaves as a monomer in solution (molecular mass of about 45 kDa). We demonstrated that SsoCdc6-1 binds single- and double-stranded DNA molecules by electrophoretic mobility shift assays. SsoCdc6-1 undergoes autophosphorylation in vitro and possesses a weak ATPase activity, whereas the protein with a mutation in the Walker A motif (Lys-59 --> Ala) is completely unable to hydrolyze ATP and does not autophosphorylate. We found that SsoCdc6-1 strongly inhibits the ATPase and DNA helicase activity of the S. solfataricus MCM protein. These findings provide the first in vitro biochemical evidence of a functional interaction between a MCM complex and a Cdc6 factor and have important implications for the understanding of the Cdc6 biological function.
- Published
- 2003
29. A preservation method that allows recovery of intact RNA from tissues dissected by laser capture microdissection
- Author
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Vincenzo Cuccurullo, Paolo Emidio Macchia, Annamaria Rosica, Luigi Mansi, Mario De Felice, J. Frederic Mushinski, James D. Owens, Rosanna Parlato, Roberto Di Lauro, Robert F. Bonner, Parlato, R, Rosica, A, Cuccurullo, Vincenzo, Mansi, Luigi, Macchia, P, Owens, Jd, Mushinski, Jf, DE FELICE, M, Bonner, Rf, DI LAURO, R., Parlato, Rosanna, Rosica, Annamaria, Macchia, PAOLO EMIDIO, Owens, J. D., Mushinski, J. F., DE FELICE, Mario, Bonner, R. F., and DI LAURO, Roberto
- Subjects
EXTRACTION ,Sucrose ,Biophysics ,Biology ,Polymerase Chain Reaction ,Biochemistry ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Histological diagnosis ,Gene expression ,Animals ,Humans ,FIXATION ,RNA, Messenger ,Thyroid Neoplasms ,Molecular Biology ,030304 developmental biology ,Laser capture microdissection ,Cryopreservation ,RNA metabolism ,Cell specific ,0303 health sciences ,cDNA library ,Dissection ,Lasers ,RNA ,AMPLIFICATION ,Cell Biology ,Blotting, Northern ,Embryo, Mammalian ,GENE ,Molecular biology ,Blot ,PCR ,030220 oncology & carcinogenesis ,Tissue Preservation - Abstract
We report a novel method for preparing samples for laser capture microdissection. The procedure described here permits extraction of intact RNA while preserving morphology, thus being suitable both for identification of specific cells and for analysis of their gene expression. The method is applicable to both mouse embryos and human tumors and may improve the preparation of cDNA libraries from specific cell types without interfering with histological diagnosis. (c)2001 Elsevier Science.
- Published
- 2002
30. Distribution of the titf2/foxe1 gene product is consistent with an important role in the development of foregut endoderm, palate and hair
- Author
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Rosanna Parlato, Roberto Di Lauro, Mario De Felice, Nina Dathan, Annamaria Rosica, Dathan, N, Parlato, R, Rosica, Am, DE FELICE, Mario, DI LAURO, Roberto, Dathan, N., Parlato, R., Rosica, A., and DE FELICE, M.
- Subjects
endocrine system ,Pharyngeal pouch ,Organogenesis ,Thyroid Gland ,030209 endocrinology & metabolism ,Ectoderm ,Biology ,Cell Line ,Mice ,03 medical and health sciences ,Esophagus ,0302 clinical medicine ,Pituitary Gland, Anterior ,Pregnancy ,medicine ,Animals ,Humans ,RNA, Messenger ,Phosphorylation ,In Situ Hybridization ,030304 developmental biology ,0303 health sciences ,Palate ,Endoderm ,Thyroid ,Embryogenesis ,Mouth Mucosa ,Forkhead Transcription Factors ,Foregut ,Anatomy ,respiratory system ,Embryo, Mammalian ,Thyroid agenesis ,Cell biology ,DNA-Binding Proteins ,Repressor Proteins ,medicine.anatomical_structure ,Female ,Digestive System ,Hair ,Transcription Factors ,Developmental Biology ,FOXE1 - Abstract
Titf2/foxe1 is a forkhead domain-containing gene expressed in the foregut, in the thyroid, and in the cranial ectoderm of the developing mouse. Titf2 null mice exhibit cleft palate and either a sublingual or completely absent thyroid gland. In humans, mutations of the gene encoding for thyroid transcription factor-2 (TTF-2) result in the Bamforth syndrome, characterized by thyroid agenesis, cleft palate, spiky hair, and choanal atresia. Here, we report a detailed expression pattern of TTF-2 protein during mouse embryogenesis and show its presence in structures where it has not been described yet. At embryonic day (E) 10.5, TTF-2 is expressed in Rathke's pouch, in thyroid, and in the epithelium of the pharyngeal wall and arches, whereas it is absent in the epithelium of the pharyngeal pouches. According to this expression, at E13.5, TTF-2 is present in endoderm derivatives, such as tongue, palate, epiglottis, pharynx, and oesophagus. Later in embryogenesis, we detect TTF-2 in the choanae and whiskers. This pattern of expression helps to define the complex phenotype displayed by human patients. Finally, we show that TTF-2 is a phosphorylated protein. These results help to characterize the domains of TTF-2 expression, from early embryogenesis throughout organogenesis, providing more detail on the potential role of TTF-2 in the development of endoderm and ectoderm derived structures.
- Published
- 2002
31. The functional ccpA gene is required for carbon catabolite repression in Lactobacillus plantarum
- Author
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Maurilio De Felice, Lidia Muscariello, Rosangela Marasco, Margherita Sacco, Muscariello, Lidia, Marasco, Rosangela, DE FELICE, M., Sacco, Margherita, Muscariello, L., Marasco, R., DE FELICE, Maurilio, and Sacco, M.
- Subjects
Dipeptidases ,Transcription, Genetic ,Molecular Sequence Data ,DNA Footprinting ,Catabolite repression ,DNA footprinting ,Repressor ,Genetics and Molecular Biology ,Biology ,Applied Microbiology and Biotechnology ,chemistry.chemical_compound ,Bacterial Proteins ,Transcription (biology) ,Gene expression ,Promoter Regions, Genetic ,Gene ,Base Sequence ,Ecology ,Promoter ,Gene Expression Regulation, Bacterial ,Molecular biology ,DNA-Binding Proteins ,Repressor Proteins ,carbohydrates (lipids) ,Lactobacillus ,chemistry ,CCPA ,bacteria ,Gene Deletion ,Food Science ,Biotechnology - Abstract
We report the characterization of the ccpA gene of Lactobacillus plantarum , coding for catabolite control protein A. The gene is linked to the pepQ gene, encoding a proline peptidase, in the order ccpA-pepQ , with the two genes transcribed in tandem from the same strand as distinct transcriptional units. Two ccpA transcription start sites corresponding to two functional promoters were found, expression from the upstream promoter being autogenously regulated through a catabolite-responsive element ( cre ) sequence overlapping the upstream +1 site. During growth on ribose, the upstream promoter showed maximal expression, while growth on glucose led to transcription from the downstream promoter. In a ccpA mutant strain, the gene was transcribed mainly from the upstream promoter in both repressing and non repressing conditions. Expression of two enzyme activities, β-glucosidase and β-galactosidase, was relieved from carbon catabolite repression in the ccpA mutant strain. In vivo footprinting analysis of the catabolite-controlled bglH gene regulatory region in the ccpA mutant strain showed loss of protection of the cre under repressing conditions.
- Published
- 2001
32. A physical and functional analysis of the newly-identified bglGPT operon of Lactobacillus plantarum
- Author
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Margherita Sacco, Rosangela Marasco, Immacolata Salatiello, Maurilio De Felice, Marasco, Rosangela, Salatiello, I., DE FELICE, M., and Sacco, Margherita
- Subjects
Operon ,Sequence analysis ,Molecular Sequence Data ,Catabolite repression ,bglGPT operon ,Biology ,medicine.disease_cause ,Microbiology ,Open Reading Frames ,Bacterial Proteins ,Genetics ,medicine ,Escherichia coli ,Cloning, Molecular ,Phosphoenolpyruvate Sugar Phosphotransferase System ,Molecular Biology ,Lactobacillu ,Base Sequence ,Permease ,beta-Glucosidase ,Membrane Transport Proteins ,RNA-Binding Proteins ,PEP group translocation ,Recombinant Proteins ,β-Glucosidase ,Complementation ,Open reading frame ,Lactobacillus ,RNA, Bacterial ,Biochemistry ,Carbon-catabolite repression ,Plasmids - Abstract
A newly-identified bglGPT operon of Lactobacillus plantarum was isolated and expressed in Escherichia coli. The sequence analysis of the cloned DNA fragment showed three open reading frames encoding (i) a 237-amino acid protein (BglG), (ii) a 577-amino acid protein (BglP) and (iii) a 486-amino acid protein (BglT). BglG, BglP and BglT were shown to be homologous to the BglG family of transcriptional antiterminators, to permeases of the phosphoenolpyruvate-dependent phosphotransferase system and to β-glucosidases, respectively. Complementation of E. coli mutant strains showed that BglP and BglT are a permease and a β-glucosidase active on the β-glucosides, 5-bromo-4-chloro-3-indolyl-β-D-glucopyranoside and p-nitrophenyl-β-D-glucoside, respectively. BglG was also shown to promote expression of a bglG-lacZ gene fusion in an E. coli bglG- background. A ribonucleic antiterminator sequence, the antiterminator-responsive cis-element and a 'catabolite responsive element', were found downstream of the transcriptional start point. Transcription of the operon was repressed 10-fold in L. plantarum cells grown on glucose as compared to ribose. Copyright (C) 2000 Federation of European Microbiological Societies.
- Published
- 2000
33. Enhanced and feedback-resistant gamma-glutamyl kinase activity of an Escherichia coli transformant carrying a mutated proBA gene of Streptococcus thermophilus
- Author
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Giuseppe Forlani, Immacolata Massarelli, Maurilio De Felice, Ezio Ricca, Massarelli, I, Forlani, G, Ricca, Ezio, DE FELICE, M., Massarelli, I., Forlani, G., Ricca, E., and DE FELICE, Maurilio
- Subjects
3-4-dehydroproline ,Streptococcus thermophilus ,Proline ,Molecular Sequence Data ,medicine.disease_cause ,Polymerase Chain Reaction ,Microbiology ,Feedback ,chemistry.chemical_compound ,Biosynthesis ,Aspartic acid ,Escherichia coli ,Genetics ,medicine ,Point Mutation ,Amino Acid Sequence ,Kinase activity ,glutamyl kinase ,proB ,proline biosynthesis ,azetidine-2-carboxylate ,Molecular Biology ,Amino acid synthesis ,chemistry.chemical_classification ,Base Sequence ,biology ,Point mutation ,Streptococcus ,Phosphotransferases (Carboxyl Group Acceptor) ,biology.organism_classification ,Amino acid ,Amino Acid Substitution ,chemistry ,Biochemistry ,Transformation, Bacterial ,Azetidinecarboxylic Acid - Abstract
We used a PCR-based method to generate a single base pair mutation in the proB gene of Streptococcus thermophilus, which replaced an aspartic acid with a glycine residue at position 192 of the first proline biosynthetic enzyme gamma-glutamyl kinase. This was the first identified mutation in amino acid biosynthesis in S. thermophilus to our knowledge. The mutation caused an enhanced, feedback-resistant gamma-glutamyl kinase activity and conferred an analogue-resistant phenotype to an Escherichia coli transformant containing the mutated gene.
- Published
- 2000
34. Expression of the bglH gene of Lactobacillus plantarum is controlled by carbon catabolite repression
- Author
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Mario Varcamonti, Lidia Muscariello, Maurilio De Felice, Margherita Sacco, Rosangela Marasco, Marasco, Rosangela, Muscariello, Lidia, Varcamonti, M., DE FELICE, M., Sacco, Margherita, Marasco, R, Muscariello, L, Varcamonti, Mario, DE FELICE, Maurilio, and Sacco, M.
- Subjects
Transcription, Genetic ,Molecular Sequence Data ,Catabolite repression ,Genetics and Molecular Biology ,Biology ,medicine.disease_cause ,Microbiology ,Enzyme Repression ,Open Reading Frames ,Glucosides ,Transcription (biology) ,Consensus Sequence ,Consensus sequence ,medicine ,Amino Acid Sequence ,RNA, Messenger ,Cloning, Molecular ,Molecular Biology ,Gene ,Escherichia coli ,Benzyl Alcohols ,Base Sequence ,Sequence Homology, Amino Acid ,biology.organism_classification ,Molecular biology ,Recombinant Proteins ,Open reading frame ,Lactobacillus ,Glucose ,Biochemistry ,Genes, Bacterial ,Sequence Alignment ,Lactobacillus plantarum ,Glucosidases - Abstract
A newly identified bglH gene coding for a phospho-β-glucosidase of Lactobacillus plantarum was isolated and expressed in Escherichia coli . The sequence analysis of the cloned DNA fragment showed an open reading frame encoding a 480-amino-acid protein with a calculated molecular mass of 53 kDa. The bglH gene was shown to be expressed on a monocistronic transcriptional unit. Its transcription was repressed 10-fold in L. plantarum cells grown on glucose compared to the β-glucoside salicin as a sole carbon source. A catabolite-responsive element (CRE) spanning from −3 to +11 with respect to the transcriptional start point was found, and its functionality was assessed by mutational analysis. In vitro and in vivo DNA binding experiments suggested the occurrence of a DNA-protein complex at the CRE site, which would mediate glucose repression of bglH expression.
- Published
- 1998
35. Transfection of TTF-1 gene induces thyroglobulin gene expression in undifferentiated FRT cells
- Author
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Gaetano Calì, Anna Mascia, Mario De Felice, Raffaele Gentile, Mariastella Zannini, Concetta Lipardi, Lucio Nitsch, Roberto Di Lauro, Mascia, A., DE FELICE, Mario, Lipardi, C., Gentile, R., Cali, G., Zannini, M., Dilauro, R., Nitsch, L., Mascia, A, DE FELICE, M, Lipardi, C, Gentile, R, Cali, G, Zannini, M, DI LAURO, R, and Nitsch, Lucio
- Subjects
endocrine system diseases ,PROMOTER ,medicine.medical_treatment ,Thyroid Nuclear Factor 1 ,Thyroid Gland ,Fluorescent Antibody Technique ,Biochemistry ,Fusion gene ,0302 clinical medicine ,TTF-1 ,thyroglobulin ,FRT cell ,transfection ,transcription factor ,Genes, Reporter ,Structural Biology ,BINDING ,Paired Box Transcription Factors ,Promoter Regions, Genetic ,TRANSGENIC MICE ,0303 health sciences ,Expression vector ,Estradiol ,Thyroid ,Nuclear Proteins ,Transfection ,respiratory system ,LRP2 ,Immunohistochemistry ,Respiratory Syncytial Viruses ,DNA-Binding Proteins ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,CHLORAMPHENICOL ACETYLTRANSFERASE ,THYROTROPIN RECEPTOR GENE ,endocrine system ,Recombinant Fusion Proteins ,Blotting, Western ,Biophysics ,Biology ,Thyroglobulin ,Cell Line ,PAX8 Transcription Factor ,03 medical and health sciences ,NUCLEAR-PROTEIN ,Genetics ,medicine ,Animals ,TRANSCRIPTION FACTOR-I ,RNA, Messenger ,Gene ,Transcription factor ,030304 developmental biology ,THYROID-CELLS ,Precipitin Tests ,Molecular biology ,Rats ,Gene Expression Regulation ,Trans-Activators ,PAX-8 ,TISSUE-SPECIFIC EXPRESSION ,Transcription Factors - Abstract
The thyroglobulin gene, the substrate for thyroid hormone biosynthesis, is not expressed in the FRT cell line, which, even though it manifests the polarised epithelial phenotype, does not express any of the thyroid functional properties. Two transcription factors, TTF-1 and Pax-8, have been implicated in thyroid specific expression of the thyroglobulin gene. FRT cells contain Pax-8 but they lack TTF-1. In this paper, we show that transfection of TTF-1 expression vectors in FRT cells results in activation of thyroglobulin gene expression. If the expression vector encoded for TTF-1-ER, a fusion gene coding for the entire TTF-1 protein fused to the hormone-binding domain of the steroid receptor, under the control of the RSV promoter, thyroglobulin gene expression was controlled by estrogen. These data provide a direct demonstration that TTF-1 activates the chromosomal thyroglobulin promoter. Since transfection of TTF-1 expression vectors in non-thyroid cell types did not result in thyroglobulin gene expression, it is suggested that Pax-8, in addition, perhaps, to a specific cellular environment, might be required for thyroid specific expression of the thyroglobulin gene. (C) 1997 Elsevier Science B.V.
- Published
- 1997
36. Membrane topology analysis of the Bacillus subtilis BofA protein involved in pro-sigma K processing
- Author
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Rosangela Marasco, Mario Varcamonti, Margherita Sacco, De Felice Maurilio, Varcamonti, Mario, Marasco, R., DE FELICE, Maurilio, Sacco, M., Varcamonti, M., Marasco, Rosangela, DE FELICE, M., and Sacco, Margherita
- Subjects
Protein Conformation ,Recombinant Fusion Proteins ,Molecular Sequence Data ,lac operon ,Bacillus subtilis ,Mutagenesi ,Microbiology ,Bacterial Proteins ,Pro-σ(K) processing ,Bacillus subtili ,Amino Acid Sequence ,Protein Precursors ,Site-directed mutagenesis ,Integral membrane protein ,Peptide sequence ,Spores, Bacterial ,Reporter gene ,biology ,fungi ,Membrane Proteins ,Alkaline Phosphatase ,biology.organism_classification ,Cell Compartmentation ,Lac Operon ,Biochemistry ,Membrane protein ,Sporulation ,Membrane topology ,Mutagenesis, Site-Directed ,Protein Processing, Post-Translational ,Gene fusion ,Transcription Factors - Abstract
The Bacillus subtilis BofA protein is involved in regulation of pro-σ K processing in the mother cell during the late stages of sporulation. A computer analysis of the BofA amino acid sequence indicates that it is an integral membrane protein. To determine the membrane topology of the protein, a series of gene fusions of bofA with lacZ or phoA reporter genes in Escherichia coli were analysed. A BofA topological model with two membrane-spanning segments, and with the N- and the C-terminal domains located in the region between the inner and outer membranes surrounding the forespore is presented. The analysis of different modifications of the last five amino acid residues of the BofA protein, obtained by PCR site-directed mutagenesis, suggests a possible role of the C-terminal domain in the regulation of pro-σ K processing.
- Published
- 1997
37. A new Bacillus subtilis gene with homology to Escherichia coli prc
- Author
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Ezio Ricca, Rosangela Marasco, Margherita Sacco, Mario Varcamonti, Marasco, R, Varcamonti, Mario, Ricca, E, DE FELICE, M. AND SACCO M., Marasco, Rosangela, Varcamonti, M., Ricca, E., and Sacco, Margherita
- Subjects
Mutant ,Molecular Sequence Data ,Sequence alignment ,Bacillus subtilis ,Biology ,medicine.disease_cause ,Homology (biology) ,Open Reading Frames ,Bacterial Proteins ,Endopeptidases ,Genetics ,medicine ,CtpA gene ,Escherichia coli ,Amino Acid Sequence ,RNA, Messenger ,Cloning, Molecular ,Promoter Regions, Genetic ,Gene ,Peptide sequence ,PBP protein ,Base Sequence ,Sequence Homology, Amino Acid ,General Medicine ,Sequence Analysis, DNA ,biology.organism_classification ,Molecular biology ,Protease ,Open reading frame ,RNA, Bacterial ,Genes, Bacterial ,Mutation ,bacteria - Abstract
We report the cloning of a 2-kb PstI-BamHI fragment of Bacillus subtilis DNA carrying an open reading frame of 1398 bp, herein designated orfRM1. This orf was shown to be transcribed only during vegetative growth from a putative sigma A-specific promoter. The deduced amino acid sequence predicted a polypeptide of 51 kDa (466 aa), which shows significant percentage of identity with the Escherichia coli Prc protein. However no Prc-like phenotypes were observed in a B. subtilis orfRM1 deletion-insertion mutant.
- Published
- 1996
38. Molecular events in the differentiation of the thyroid gland
- Author
-
Mariastella Zannini, Giuseppe Damante, Koichi Sato, Renata Lonigro, Maria Ina Arnone, M. De Felice, Roberto Di Lauro, DI LAURO, Roberto, Damante, G, DE FELICE, Mario, Arnone, Mi, Sato, K, Lonigro, R, Zannini, M., DI LAURO, R., Damante, G., DE FELICE, M., Arnone, M. I., Sato, K., Lonigro, R., and Zannini, Mariastella
- Subjects
medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,Thyroid Nuclear Factor 1 ,Thyroid Gland ,030209 endocrinology & metabolism ,Biology ,Iodide Peroxidase ,Thyroglobulin ,03 medical and health sciences ,PAX8 Transcription Factor ,0302 clinical medicine ,Endocrinology ,Thyroid peroxidase ,Transcription (biology) ,Internal medicine ,medicine ,Animals ,Paired Box Transcription Factors ,RNA, Messenger ,Promoter Regions, Genetic ,030304 developmental biology ,Homeodomain Proteins ,0303 health sciences ,Thyroid ,Nuclear Proteins ,Cell Differentiation ,Cell biology ,Rats ,DNA-Binding Proteins ,Gene Expression Regulation, Neoplastic ,medicine.anatomical_structure ,biology.protein ,Trans-Activators ,Homeobox ,PAX8 ,Transcription Factors - Published
- 1995
39. Utilization of cellobiose and other β-D-glucosides in Agrobacterium tumefaciens
- Author
-
M. De Felice, Rosangela Marasco, C. T. Lago, Marasco, Rosangela, and Lago, C. T. AND DE FELICE M.
- Subjects
Cellobiose, Catabolite repression, β-Glucosidase, Agrobacterium tumefacien ,Cellobiose ,Rhizobiaceae ,Catabolite repression ,In Vitro Techniques ,β-D-glucosides, Cyclic AMP, Virulence ,Microbiology ,chemistry.chemical_compound ,Hydrolysis ,Glucosides ,Glucoside ,Cyclic AMP ,Molecular Biology ,chemistry.chemical_classification ,biology ,beta-Glucosidase ,General Medicine ,Metabolism ,Agrobacterium tumefaciens ,biology.organism_classification ,Glucose ,Enzyme ,chemistry ,Biochemistry ,Depression, Chemical ,Electrophoresis, Polyacrylamide Gel - Abstract
Agrobacterium tumefaciens strain C58 was able to utilize carbon from cellobiose and some other beta-D-glucosides as efficiently as from glucose. beta-D-glucoside utilization was partially inducible and the induction was subject to catabolite repression by glucose, independently of the presence of cyclic AMP in the medium. It was also independent of Ti plasmid-encoded functions. beta-D-glucosides were hydrolysed by a single, cytoplasmic and constitutively expressed beta-glucosidase, which was active on non-phosphorylated substrates and insensitive to glucose inhibition.
- Published
- 1995
40. Control of ilvIH transcription during amino acid downshift in stringent and relaxed strain of Escherichia coli
- Author
-
Maurilio De Felice, Margherita Sacco, Loredana Baccigalupi, Rosangela Marasco, Ezio Ricca, Baccigalupi, L, Marasco, R, Ricca, E, DE FELICE, Maurilio, Sacco, M., DE FELICE, M, Sacco, Margherita, and Ricca, Ezio
- Subjects
Transcription, Genetic ,Stringent response ,Operon ,Molecular Sequence Data ,medicine.disease_cause ,Microbiology ,Bacterial Proteins ,Transcription (biology) ,relA ,Genetics ,medicine ,Escherichia coli ,Amino Acids ,Molecular Biology ,chemistry.chemical_classification ,Messenger RNA ,ilvIH transcription ,biology ,Base Sequence ,Escherichia coli Proteins ,Gene Expression Regulation, Bacterial ,biology.organism_classification ,Enterobacteriaceae ,Molecular biology ,Leucine-Responsive Regulatory Protein ,lrp transcription ,Amino acid ,DNA-Binding Proteins ,RNA, Bacterial ,Biochemistry ,chemistry ,Genes, Bacterial ,bacteria ,Bacteria ,Transcription Factors - Abstract
Transcription of the ilvIH operon was reduced during amino acid starvation of wild-type Escherichia coli. The effect was abolished by a relA mutation and was enhanced by a spoT mutation, thus suggesting a possible negative role of ppGpp accumulation on ilvIH transcription. No effect of amino acid downshift was observed on the synthesis of Irp mRNA, encoding the positive regulator (Lrp) of ilvIH transcription. © 1995.
- Published
- 1995
41. Acetohydroxy acid synthase and threonine deaminase activities, and the biosynthesis of isoleucine-leucine-valine in Streptococcus bovis
- Author
-
C Caruso, M. De Felice, A L Basso, Ezio Ricca, L Ferrara, Basso, Al, Ricca, E, Caruso, C, Ferrara, L, DE FELICE, Maurilio, Ricca, Ezio, and DE FELICE, M.
- Subjects
chemistry.chemical_classification ,Acetolactate synthase ,biology ,Threonine Dehydratase ,Valine ,General Medicine ,Microbiology ,Amino acid ,Streptococcus bovis ,chemistry.chemical_compound ,Acetolactate Synthase ,Butyrates ,chemistry ,Biosynthesis ,Biochemistry ,Leucine ,biology.protein ,Isoleucine ,Threonine ,Molecular Biology - Abstract
Acetohydroxy acid synthase (AHAS) and threonine deaminase (TD) activities were found in Streptococcus bovis and shown to be involved in the biosynthesis of the branched chain amino acids isoleucine, leucine and valine. Apparent lack of repression of AHAS synthesis by the end-products and reduced sensitivity of S. bovis growth to analogues of the branched chain amino acids suggested that secretion of isoleucine, leucine and valine in the growth medium may be a consequence of the regulatory features of AHAS. A glycyl-leucine-resistant mutant with reduced TD activity secreted a reduced amount of isoleucine and an increased amount of valine, which might be a result of the reduced rate of synthesis of the isoleucine precursor alpha-ketobutyrate and of a consequent preferential carbon flow through the valine branch of the pathway.
- Published
- 1993
42. A stereospecific alignment between the promoter and the cis-acting sequence is required for Lrp-dependent activation of ilvIH transcription in Escherichia coli
- Author
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Rosangela Marasco, Roberta Paradiso, Ezio Ricca, Maurilio De Felice, Margherita Sacco, Sacco, Margherita, Ricca, E, Marasco, Rosangela, and Paradiso, R. AND DE FELICE M.
- Subjects
DNA, Bacterial ,ilvIH ,Transcription, Genetic ,Operon ,Lrp ,Molecular Sequence Data ,DNA-binding protein ,Biology ,Microbiology ,Protein-protein interaction ,Facing ,Bacterial Proteins ,Transcription (biology) ,Genetics ,Escherichia coli ,Transcription activation ,Insertion ,Promoter Regions, Genetic ,Molecular Biology ,Gene ,Regulation of gene expression ,Base Sequence ,Escherichia coli Proteins ,Gene Expression Regulation, Bacterial ,Cis-Acting Sequence ,Molecular biology ,Leucine-Responsive Regulatory Protein ,DNA-Binding Proteins ,Regulatory sequence ,Genes, Bacterial ,Mutagenesis ,lipids (amino acids, peptides, and proteins) ,Plasmids ,Transcription Factors - Abstract
The leucine-responsive regulatory protein (Lrp) is a DNA binding protein that affects, either positively or negatively, the expression of several E. coli genes. The ilvIH operon is positively regulated by Lrp and leucine counteracts this effect reducing 5- to 10-fold the efficiency of ilvIH transcription. An investigation of the mechanism of transcription activation of the ilvIH operon by Lrp indicated that: (i) a stereospecific alignment between the ilvIH promoter and the cis-acting sequence upstream of it is required for activation; (ii) a correct distance between the promoter and the adjacent cis-acting sequence is needed for leucine to counteract the positive role of Lrp; (iii) Lrp fails to activate transcription when the cis-acting region is placed several hundred base pairs upstream of the ilvIH promoter. © 1993.
- Published
- 1993
43. ORGANIZATION OF LRP-BINDING SITES UPSTREAM OF ILVIH IN SALMONELLA-TYPHIMURIUM
- Author
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Ezio Ricca, Margherita Sacco, C. T. Lago, Joseph M. Calvo, Qing Wang, Maurilio DeFelice, Wang, Q, Sacco, Margherita, Ricca, E, Lago, Ct, DE FELICE, M, and Calvo, M.
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DNA, Bacterial ,Salmonella typhimurium ,Operon ,Molecular Sequence Data ,medicine.disease_cause ,Microbiology ,Bacterial Proteins ,Species Specificity ,Sequence Homology, Nucleic Acid ,Consensus Sequence ,Escherichia coli ,Leucine-responsive regulatory protein ,medicine ,Consensus sequence ,Binding site ,Promoter Regions, Genetic ,Molecular Biology ,Binding Sites ,Base Sequence ,biology ,Escherichia coli Proteins ,Nucleic acid sequence ,biology.organism_classification ,Leucine-Responsive Regulatory Protein ,Enterobacteriaceae ,Molecular biology ,Footprinting ,DNA-Binding Proteins ,Acetolactate Synthase ,Genes, Bacterial ,biology.protein ,bacteria ,lipids (amino acids, peptides, and proteins) ,Sequence Alignment ,Transcription Factors - Abstract
Lrp, a major regulatory protein in Escherichia coli, controls the expression of numerous operons, including ilvlH. Lrp binds to six sites upstream of ilvlH, and Lrp binding is required for ilvlH expression. We show here that an Lrp‐like protein is also present in Salmonella typhimurium. This protein can bind both E. coli and S. typhimurium ilvlH DNA, as can E. coli Lrp. Methidiumpropyl‐EDTA footprinting studies were performed with purified E. coli Lrp and S. typhimurium ilvlH DNA. Six binding sites were defined, three of them being similar to corresponding sites in E. coli, and three being organized differently. A consensus derived from six S. typhimurium sites is compatible with that derived from a similar analysis of E. coli sequences. Copyright © 1993, Wiley Blackwell. All rights reserved
- Published
- 1993
44. Absence of acetohydroxy acid synthase III in Salmonella typhimurium is due to early termination of translation within the ilvl gene
- Author
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C. T. Lago, Margherita Sacco, Ezio Ricca, M. Fellce, Ricca, E, Lago, Ct, Sacco, Margherita, DE FELICE, M., Sacco, M, DE FELICE, Maurilio, and Ricca, Ezio
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Genetics ,Salmonella typhimurium ,Base Sequence ,Operon ,Protein subunit ,Mutant ,Molecular Sequence Data ,Nucleic acid sequence ,Locus (genetics) ,Biology ,Peptide Chain Termination, Translational ,Regulatory Sequences, Nucleic Acid ,Microbiology ,Molecular biology ,Isoenzymes ,Acetolactate Synthase ,Genes, Bacterial ,Coding region ,Transversion ,Molecular Biology ,Gene - Abstract
The cryptic ilvlH locus of Salmonella typhimurium has genetic information for two distinct subunits of acetohydroxy acid synthase III. We show that the ilvH‐encoded subunit is normally translated and the lack of activity is due to early termination of translation within the promoter‐proximal ilvl gene. Analysis of the 5’region of the operon led to identification of the promoter and the amino‐terminal part of ilvl. Expression of this gene in a mutant producing acetohydroxy acid synthase is due to a transversion which creates a UUA (leucine) codon in the place of a UGA (stop) codon present in position 12 of the wild‐type coding region. Copyright © 1991, Wiley Blackwell. All rights reserved
- Published
- 1991
45. Mitogenic Activity of Anti-cd28 Moab Clb-cd28/1 On Peripheral-blood Mononuclear-cells and Its Cooperation With Other Anti-t Cells Moab In the Activation of Purified Lymphocytes-t
- Author
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Patrizia Carandente Giarrusso, R. A. W. Van Lier, Salvatore Venuta, Soldano Ferrone, Giuliana Valerio, Soo Young Yang, Maria Caterina Turco, M De Felice, Annalisa Lamberti, DE FELICE, Mario, P. C., Giarrusso, Lamberti, Annalisa, M. C., Turco, G., Valerio, R. A., W., S. Y., Yang, S., Ferrone, S., Venuta, De Felice, M., Giarrusso, P. C., Turco, M. C., Valerio, G., van Lier, R., Yang, S. Y., Ferrone, S., Venuta, S., and Other departments
- Subjects
Antigens, Differentiation, T-Lymphocyte ,Interleukin 2 ,CD3 Complex ,T-Lymphocytes ,CD3 ,Immunology ,Receptors, Antigen, T-Cell ,chemical and pharmacologic phenomena ,Thymus Gland ,CD5 Antigens ,Lymphocyte Activation ,Biochemistry ,Peripheral blood mononuclear cell ,Monocytes ,Immunoglobulin Fab Fragments ,CD28 Antigens ,Genetics ,medicine ,Humans ,Immunology and Allergy ,Antigen-presenting cell ,biology ,Interleukins ,Monocyte ,Histocompatibility Antigens Class I ,Antibodies, Monoclonal ,CD28 ,hemic and immune systems ,General Medicine ,T lymphocyte ,Antigens, Differentiation ,Molecular biology ,medicine.anatomical_structure ,Leukocytes, Mononuclear ,biology.protein ,CD5 ,Cell Division ,medicine.drug - Abstract
Human T lymphocyte proliferative response induced via CD28 molecule is analyzed. An anti CD28 MoAb, CLB-CD28/1, induces the proliferation of human peripheral blood mononuclear cells in the absence of other stimuli, indicating that CD28 molecule can directly mediate a mitogenic signal in this system. The mitogenic activity of MoAb CLB-CD28/1 on PBMC does not require MoAb interaction with monocyte Fc receptors, since F(ab′)2 fragments from the MoAb are mitogenic to the same extent as whole IgG. Nevertheless, the activity depends on the presence of accessory cells, since purified T lymphocytes require addition of irradiated monocytes and interleukin 2 to proliferate when incubated with MoAb CLB-CD28/1. On the other hand, MoAb CLB-CD28/1 induces response to IL-2 in thymocytes in the absence of accessory cells. Cooperation of MoAb CLB-CD28/1 with three other MoAbs, recognizing CD3, CD5 and HLA Class I antigens, respectively, induces Tac antigen expression and IL-2 responsiveness in purified T lymphocytes. This effect is obtained without cross-linking of the MoAb. It does not rely on a physical association between CD28 and CD3, CD5 or HLA Class I molecules, as demonstrated by co-modulation experiments. These data indicate that expression of IL-2 receptor on T lymphocytes can result from interaction of multiple activation pathways and that some of them, such as those mediated by CD5 and HLA Class I antigens, previously reported to serve as modulatory circuits, can instead act as essential elements in the onset of T lymphocyte proliferation.
- Published
- 1990
46. Antimicrobial activity of a newly identified bacteriocin of Bacillus cereus
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Ezio Ricca, M Sacco, M De Felice, G Naclerio, Naclerio, G, Ricca, E, Sacco, Margherita, DE FELICE, M., Sacco, M, DE FELICE, Maurilio, and Ricca, Ezio
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Bacillaceae ,Ecology ,biology ,Molecular mass ,Bacillus cereus ,biology.organism_classification ,Antimicrobial ,Applied Microbiology and Biotechnology ,Bacillales ,Microbiology ,Cereus ,Biochemistry ,Bacteriocin ,Bacteriocins ,bacteria ,Bacteria ,Food Science ,Biotechnology ,Research Article - Abstract
A bacteriocin-producing Bacillus cereus strain was isolated. The bacteriocin, here called cerein, was shown to be active specifically against other B. cereus strains and inactive against all other bacterial species tested. Cerein was detected in the culture supernatants of stationary-phase cells, and its appearance was inhibited by induction of sporulation. The bacterial activity of cerein was insensitive to organic solvents and nonproteolytic enzymes, partially stable to heat, and active over a wide range of pH values. Direct detection of antimicrobial activity on sodium dodecyl sulfate-polyacrylamide gel suggested an apparent molecular mass of about 9 kDa.
47. Bacillus subtilis spore coat assembly requires cotH gene expression
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M De Felice, Loredana Baccigalupi, Ezio Ricca, Gino Naclerio, Rita Zilhão, Naclerio, G, Baccigalupi, L., Zilhao, R, DE FELICE, Maurilio, Ricca, E., Baccigalupi, L, DE FELICE, M, and Ricca, Ezio
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DNA, Bacterial ,Mutant ,Molecular Sequence Data ,Gene Expression ,Bacillus subtilis ,Biology ,Microbiology ,Endospore ,Open Reading Frames ,Bacterial Proteins ,Gene expression ,Amino Acid Sequence ,RNA, Messenger ,Molecular Biology ,Gene ,Peptide sequence ,Spores, Bacterial ,Base Sequence ,fungi ,Correction ,biology.organism_classification ,Molecular biology ,Spore ,Cell biology ,Open reading frame ,Genes, Bacterial ,Mutation ,Research Article - Abstract
Endospores of Bacillus subtilis are encased in a protein shell, known as the spore coat, composed of a lamella-like inner layer and an electron-dense outer layer. We report the identification and characterization of a gene, herein called cotH, located at 300 degrees on the B. subtilis genetic map between two divergent cot genes, cotB and cotG. The cotH open reading frame extended for 1,086 bp and corresponded to a polypeptide of 42.8 kDa. Spores of a cotH null mutant were normally heat, lysozyme, and chloroform resistant but were impaired in germination. The mutant spores were also pleiotropically deficient in several coat proteins, including the products of the previously cloned cotB, -C, and -G genes. On the basis of the analysis of a cotE cotH double mutant, we infer that CotH is probably localized in the inner coat and is involved in the assembly of several proteins in the outer layer of the coat.
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