31 results on '"De Felice, M."'
Search Results
2. Evaluation of the Molecular Landscape in PD-L1 Positive Metastatic NSCLC: Data from Campania, Italy
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Pasquale Pisapia, Antonino Iaccarino, Caterina De Luca, Gennaro Acanfora, Claudio Bellevicine, Roberto Bianco, Bruno Daniele, Luisa Ciampi, Marco De Felice, Teresa Fabozzi, Luigi Formisano, Pasqualina Giordano, Cesare Gridelli, Giovanni Pietro Ianniello, Annamaria Libroia, Paolo Maione, Mariantonia Nacchio, Fabio Pagni, Giovanna Palmieri, Francesco Pepe, Gianluca Russo, Maria Salatiello, Antonio Santaniello, Rachele Scamarcio, Davide Seminati, Michele Troia, Giancarlo Troncone, Elena Vigliar, Umberto Malapelle, Pisapia, P, Iaccarino, A, De Luca, C, Acanfora, G, Bellevicine, C, Bianco, R, Daniele, B, Ciampi, L, De Felice, M, Fabozzi, T, Formisano, L, Giordano, P, Gridelli, C, Ianniello, G, Libroia, A, Maione, P, Nacchio, M, Pagni, F, Palmieri, G, Pepe, F, Russo, G, Salatiello, M, Santaniello, A, Scamarcio, R, Seminati, D, Troia, M, Troncone, G, Vigliar, E, Malapelle, U, Pisapia, Pasquale, Iaccarino, Antonino, DE LUCA, Caterina, Acanfora, Gennaro, Bellevicine, Claudio, Bianco, Roberto, Daniele, Bruno, Ciampi, Luisa, De Felice, Marco, Fabozzi, Teresa, Formisano, Luigi, Giordano, Pasqualina, Gridelli, Cesare, Pietro Ianniello, Giovanni, Libroia, Annamaria, Maione, Paolo, Nacchio, Mariantonia, Pagni, Fabio, Palmieri, Giovanna, Pepe, Francesco, Russo, Gianluca, Salatiello, Maria, Santaniello, Antonio, Scamarcio, Rachele, Seminati, Davide, Troia, Michele, Troncone, Giancarlo, Vigliar, Elena, and Malapelle, Umberto
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molecular oncology ,molecular pathology ,PD-L1 ,immune-checkpoint inhibitors ,biomarkers ,immune-checkpoint inhibitor ,Lung Neoplasms ,Organic Chemistry ,General Medicine ,Protein-Tyrosine Kinases ,Catalysis ,B7-H1 Antigen ,Computer Science Applications ,Inorganic Chemistry ,Proto-Oncogene Proteins p21(ras) ,Carcinoma, Non-Small-Cell Lung ,Proto-Oncogene Proteins ,Mutation ,Humans ,biomarker ,Physical and Theoretical Chemistry ,Molecular Biology ,Spectroscopy ,Retrospective Studies - Abstract
Background: Immune-checkpoint inhibitors (ICIs) have increased and improved the treatment options for patients with non-oncogene-addicted advanced stage non-small cell lung cancer (NSCLC). However, the role of ICIs in oncogene-addicted advanced stage NSCLC patients is still debated. In this study, in an attempt to fill in the informational gap on the effect of ICIs on other driver mutations, we set out to provide a molecular landscape of clinically relevant oncogenic drivers in programmed death-ligand 1 (PD-L1) positive NSCLC patients. Methods: We retrospectively reviewed data on 167 advanced stage NSCLC PD-L1 positive patients (≥1%) who were referred to our clinic for molecular evaluation of five driver oncogenes, namely, EGFR, KRAS, BRAF, ALK and ROS1. Results: Interestingly, n = 93 (55.7%) patients showed at least one genomic alteration within the tested genes. Furthermore, analyzing a subset of patients with PD-L1 tumor proportion score (TPS) ≥ 50% and concomitant gene alterations (n = 8), we found that n = 3 (37.5%) of these patients feature clinical benefit with ICIs administration, despite the presence of a concomitant KRAS gene alteration. Conclusions: In this study, we provide a molecular landscape of clinically relevant biomarkers in NSCLC PD-L1 positive patients, along with data evidencing the clinical benefit of ICIs in patient NSCLC PD-L1 positive alterations.
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- 2022
3. Novel Therapeutic Opportunities in Neoadjuvant Setting in Urothelial Cancers: A New Horizon Opened by Molecular Classification and Immune Checkpoint Inhibitors
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Maria Lucia Iacovino, Chiara Carmen Miceli, Marco De Felice, Biagio Barone, Luca Pompella, Francesco Chiancone, Erika Di Zazzo, Giuseppe Tirino, Carminia Maria Della Corte, Ciro Imbimbo, Ferdinando De Vita, Felice Crocetto, Iacovino, M. L., Miceli, C. C., De Felice, M., Barone, B., Pompella, L., Chiancone, F., Di Zazzo, E., Tirino, G., Della Corte, C. M., Imbimbo, C., De Vita, F., and Crocetto, F.
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QH301-705.5 ,Immune Checkpoint Inhibitor ,Urothelial cancers ,Cystectomy ,Neoadjuvant chemotherapy ,Catalysis ,Inorganic Chemistry ,Molecular classification of muscle-invasive bladder cancer ,Antineoplastic Combined Chemotherapy Protocols ,Urothelial cancer ,Biomarkers, Tumor ,Humans ,Biology (General) ,Physical and Theoretical Chemistry ,QD1-999 ,Molecular Biology ,Immune Checkpoint Inhibitors ,Spectroscopy ,Carcinoma, Transitional Cell ,Clinical Trials as Topic ,Antineoplastic Combined Chemotherapy Protocol ,Organic Chemistry ,Bladder cancer ,General Medicine ,Neoadjuvant Therapy ,Computer Science Applications ,Gene Expression Regulation, Neoplastic ,Chemistry ,Urinary Bladder Neoplasms ,Lymph Node Excision ,Human ,Muscle-invasive bladder cancer - Abstract
Muscle invasive bladder cancer (MIBC) is a widespread malignancy with a worse prognosis often related to a late diagnosis. For early-stage MIBC pts, a multidisciplinary approach is mandatory to evaluate the timing of neoadjuvant chemotherapy (NAC) and surgery. The current standard therapy is platinum-based NAC (MVAC-methotrexate, vinblastine, doxorubicin, and cisplatin or Platinum–Gemcitabine regimens) followed by radical cystectomy (RC) with lymphadenectomy. However, preliminary data from Vesper trial highlighted that dose-dense NAC MVAC is endowed with a good pathological response but shows low tolerability. In the last few years, translational-based research approaches have identified several candidate biomarkers of NAC esponsiveness, such as ERCC2, ERBB2, or DNA damage response (DDR) gene alterations. Moreover, the recent consensus MIBC molecular classification identified six molecular subtypes, characterized by different sensitivity to chemo- or targeted or immunotherapy, that could open a novel procedure for patient selection and also for neoadjuvant therapies. The Italian PURE-01 phase II Trial extended data on efficacy and resistance to Immune Checkpoint Inhibitors (ICIs) in this setting. In this review, we summarize the most relevant literature data supporting NAC use in MIBC, focusing on novel therapeutic strategies such as immunotherapy, considering the better patient stratification and selection emerging from novel molecular classification.
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- 2021
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. 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
7. 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
8. 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
9. 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
10. 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
11. '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
12. 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
13. 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
14. 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.
- Published
- 2007
15. 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
16. 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
17. 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
18. 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
19. 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
20. 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
21. 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
22. 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
23. 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
24. Utilization of cellobiose and other β-D-glucosides in Agrobacterium tumefaciens
- Author
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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
25. Control of ilvIH transcription during amino acid downshift in stringent and relaxed strain of Escherichia coli
- Author
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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
26. Acetohydroxy acid synthase and threonine deaminase activities, and the biosynthesis of isoleucine-leucine-valine in Streptococcus bovis
- Author
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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
27. 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
28. 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.
- Subjects
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
29. 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
- Subjects
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
30. 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.
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- 1990
31. Bacillus subtilis spore coat assembly requires cotH gene expression
- Author
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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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