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Abstract: Here, we report the identification of a novel domain – GG (domain in KIAA1199, FAM3, POMGnT1 and Tmem2 proteins, with two well-conserved glycine residues), present in eukaryotic FAM3 superfamily (FAM3A, FAM3B, FAM3C and FAM3D), POMGnT1 (protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase), TEM2 proteins as well as phage gp35 proteins. GG domain has been revealed to be implicated in muscle–eye–brain disease and non-syndromic hearing loss. The presence of GG domain in Bacteriophage gp35 hinge connector of long tail fiber might reflect the horizontal gene transfer from organisms. And we proposed that GG domain might function as important structural element in phage LTF. [Copyright &y& Elsevier]

Investigates the phylogeny of primary-symbionts of species belonging to the subgeneral of Glossina based on 16S recombinant DNA sequences. Lineage of organisms in the gamma-subdivision of proteobacteria; Association of evolution with symbiosis; Evidence for the development of symbiosis acquisition by S-endosymbionts.

Abstract: Present work reported a novel domain – D8C (domain with conserved eight cysteines in liver-specific ZP domain-containing protein, glycoprotein 2 (GP-2) and uromodulin (UMOD)), present in liver-specific LZP, UMOD, GP-2 and some uncharacterized proteins, most of which are membrane proteins, extracellular proteins or nuclear membrane proteins. D8C contains eight well-conserved cysteine residues, which were predicted to form four pairs of disulfide bridges. D8C is composed mainly of β-strands. Mutation in the D8C at Cys217 in human UMOD is associated with familial juvenile hyperuricaemic nephropathy, which might be due to the disruption of the disulfide bridge. Identification of D8C would further the understandings of related proteins. [Copyright &y& Elsevier]

Abstract: Cyclophilin A (CypA), a peptidyl–prolyl cis–trans isomerase (PPIase), was originally identified as the intracellular receptor for cyclosporin A (CsA). Recently, correlations of CypA with tumor pathogenesis have been studied. Here, we studied the expression of CypA and its receptor CD147 in several kinds of lung cancer cells as well as a normal lung cell and found that in H446 cell, a kind of small cell lung cancer cell, the expression are the highest. The exogeneous CypA protein can substantially stimulate H446 cell growth in dependence on its PPIase activity. We also showed that CypA protein can stimulate ERK1/2 signal in dose and time dependent manners and almost has no effect to p38 and JNK signals. Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for small cell lung cancer. [Copyright &y& Elsevier]

Abstract: Aurora-A is a centrosome-localized serine/threonine kinase that is overexpressed in multiple human cancers. Here, we report an intramolecular inhibitory regulation in Aurora-A between its N-terminal regulatory domain (aa 1–128, Nt) and the C-terminal catalytic domain (aa 129–403, Cd). Removal of Nt results in a significant increase in kinase activity. Nt inhibited the activity of the single C-terminal kinase domain, but had little effect on the activity of the full-length of Aurora-A. PP1 is not involved in this regulation, instead, Nt interacts Cd directly in vitro and in vivo. The non-Aurora box (aa 64–128) in the N-terminal negatively regulated the kinase activity of the C-terminal kinase domain by intramolecular interaction with aa 240–300 within the C-terminal. [Copyright &y& Elsevier]

Abstract: Several families of kinases work together to ensure the rate and precision of mitosis. Aurora-B is an important serine/threonine kinase required for chromosome segregation and cytokinesis. Identification of Aurora-B substrates will help to enhance our understanding of the molecular mechanism of mitosis. Through a yeast two-hybrid screen, we found a novel partner of Aurora-B, Septin1, belonging to a conserved family of GTPase proteins that localize to the cleavage furrow and are involved in cytokinesis. We confirmed this interaction using Co-immunoprecipitation experiments in mammalian cells and GST-pull-down analysis in vitro. Moreover, Aurora-B can phosphorylate Septin1 in vitro. We identified that Ser248, Ser307, and Ser315 are the main phosphorylation sites in Septin1. These two proteins partially co-localize to the midbody during cytokinesis. So, it is possible that Septin1’s role in the regulation of cytokinesis is related to its phosphorylation by Aurora-B. Unlike previous reports that Septins function in cytokinesis and localize to the cleavage furrow, we found that Septin1 localizes to the spindle pole throughout mitosis, indicating that Septin1 may function in chromosome segregation as well. [Copyright &y& Elsevier]

Abstract: Cellular adhesion plays important roles in a variety of biological processes. The ADAM family contains disintegrin-like and metalloproteinase-like domains which potentially have cell adhesion and protease activities. Recent studies suggest that the interaction between 14-3-3ζ and ADAM22cyt can regulate cell adhesion and spreading, therefore it has a potential role in neural development and function. 14-3-3 family has seven highly conserved members that regulate various cellular functions. Using yeast two-hybrid method, we identified that ADAM22cyt bound some other 14-3-3 family members. The interaction was further confirmed by in vitro protein pull-down assay and co-immunoprecipitation. We also found that the overexpression of exogenous ADAM22 in HEK293 cells could significantly enhance cell adhesion and spreading, compared with the truncated ADAM22 lack of 14-3-3 binding motifs. These results strongly demonstrated a functional role for ADAM22/14-3-3 in cell adhesion and spreading. [Copyright &y& Elsevier]

The present work reported the cloning and characterization of two novel human genes—HN1 (hematopoietic- and neurologic-expressed sequence 1) and HN1L (HN1-like gene) which are proposed to be involved in embryo development. HN1 is mapped on chromosome 17q25.2, with two transcripts (1.0 and 1.6 kb in length, respectively) due to alternative splicing. HN1 is expressed abundantly in testis and skeletal muscle among 16 human tissues, and it is localized in the nucleus indicated by GFP fusion expression. Western blot confirmed that HN1 encodes a 16.5-kDa protein. HN1L is on chromosome 16p13.3, with three splicing in the length of 2.0, 4.0 and 4.2 kb, respectively. HN1L is expressed in a variety of tissues such as liver, kidney, prostate, testis and uterus at varying levels. HN1L gene encodes a 20-kDa protein, which is localized in both the nucleus and cytoplasm. Fourteen of HN1 and sixteen of HN1L homologous genes in different species were determined and analyzed by BLAST searches. Silicon analyses of the 14 orthologous proteins of HN1 and 16 orthologous proteins of HN1L revealed that they share great conservation in vertebrate. Additionally, we identified nine pseudogenes of HN1 (six) and HN1L (three) in the genomes of the human, mouse and rat. Based on sequence alignments and phylogenetic analysis, all these homologous genes and pseudogenes were defined as a HN1 gene family. [Copyright &y& Elsevier]

A metalloprotease and disintegrin (ADAM) is a family of membrane-anchored proteins and all family members have a multi-domain structure containing a zinc metalloprotease domain and a disintegrin domain that may serve as an integrin ligand. Here we reported two novel mammalian transcripts of Adam23, named Adam23β and Adam23γ, to be involved in the development and functional activities of mammalian brains. Adam23γ was isolated from a 22-week human fetal brain cDNA library, using an EST homologous to Adam as a probe, and is 100% homologous to human Adam23 (Adam23α) except that it lacks a fragment of 91 bp near the C-terminal, thus it could not form obvious transmembrane domain. Adam23β was discovered while the diversity at the transmembrane domain (TM) was analyzed. Adam23β has a different sequence in the 91 nucleotides and thus encode different transmembrane domain. Adam23β and Adam23γ are mainly expressed in brain like Adam23α. RT-PCR experiments in mouse brain also detected the two isoforms, consistent with observation of Northern analysis of human RNAs. Furthermore, results of RT-PCR amplification of Adam23γ in mouse brains of different developmental stages revealed a developmentally regulated expression pattern: Adam23γ is expressed in embryonic and infant brain, and disappeared after the 10th postnatal day. This temporally changing expression pattern of Adam23γ suggests that ADAM23γ likely plays an important role in brain development. [Copyright &y& Elsevier]

bri3 was identified to be a novel gene up-regulated in TNF-treated cells with suppressed subtractive hybridization (SSH) in our laboratory. Previous studies showed that overexpression of BRI3 induced apoptosis in L929 cells. To further study the function of bri3, we disrupted its expression by expressing bri3 antisense RNA. The antisense RNA promoted resistance to TNF-induced cell death by more than 1000-fold in L929 cells, suggesting the involvement of BRI3 in TNF-induced cell death in this cell line. Analysis of cell death caused by other apoptotic inducers showed that the effect of BRI3 antisense RNA is highly specific to TNF-induced cell death. Taken together, bri3 appears to play an important role in TNF-induced cell death. Finally, we reported here that BRI3 may be localized to lysosome and function through lysosome. [Copyright &y& Elsevier]

The incidence of hepatoma is high in the Chinese population. Searching for genes involved in the functions of the liver, especially genes specifically expressed in the liver, will facilitate an insight into the molecular basis of normal and abnormal liver functions. Based on a differentially displayed cDNA fragment, which was down regulated in hepatoma tissues, we cloned a novel cDNA of 957 bp, TCP10L (T-complex protein 10 like), from the human liver cDNA library. Northern hybridization of this novel gene in 30 adult human tissues was examined. The result revealed that TCP10L expressed specifically in the human liver and testis. The TCP10L contains a 645-bp open reading frame encoding a deduced protein of 215 amino acids. As the deduced protein was analyzed further, a typical leucine zipper motif was found. We firstly examined the transcriptional function of the TCP10L protein by transfecting recombinant pM-TCP10L into mammalian cells. The subsequent analysis based on the dual luciferase assay system showed that TCP10L significantly inhibited the expression of reporter genes. Compared with that of the negative control, the luciferase activity were down regulated in HEK293 and SK-HEP-1, CHO cells by about 2.6, 9.8, and 5.5 folds respectively. A mutated type of TCP10L was also constructed. It showed that the repression of TCP10L to the expression of the reporter gene almost completely decreased, suggesting that the leucine zipper structure is critical for TCP10L to play its role in regulation function. Then we transfected the recombinant TCP10L-EGFP into cells. The results indicated that TCP10L subcellularly located in nuclei, either in HEK 293 or SK-HEP-1 cells. In addition, human TCP10L was found comprised of five exons and four introns, and mapped to chromosome 21q22.11. [ABSTRACT FROM AUTHOR]

We have identified two novel human SCAN domain genes ZNF396 and ZNF397, which are clustered within the region of chromosome 18q12. Three isoforms of ZNF396 transcript, 1.5, 2.5 and 3.9-kb, are expressed highly in liver. Four isoforms of ZNF397 transcript, 1.7, 2.5, 7.0 and 9.0-kb, are expressed in a variety of tissues, with varying levels. The SCAN-(C2H2)X genes encode two distinct proteins due to a unique alternative splicing mechanism. Both ZNF396-fu (full zinc fingers) and ZNF397-fu consist of a SCAN domain in the N-terminal region and many consecutive C2H2 zinc finger repeats in the C-terminal region. ZNF396-nf (no zinc fingers) and ZNF397-nf encode 210 and 198 amino acids, respectively, containing the SCAN domain only. ZNF396-fu, ZNF396-nf, ZNF397-fu or ZNF397-nf can homo-associate, while ZNF396-fu hetero-associates with ZNF396-nf, and ZNF397-fu hetero-associates with ZNF397-nf. ZNF396-nf and ZNF397-nf polypeptides are expressed diffusely in the cells, while ZNF396-fu and ZNF397-fu polypeptides target specifically to the nuclei. ZNF396-fu, ZNF396-nf and ZNF397-nf can repress reporter gene transcription, with ZNF397-nf having the strongest repression activity. Deletion analysis revealed that ZNF397-fu is a transcriptional activator without its nine zinc finger repeats. [Copyright &y& Elsevier]

The ADAM family consists of a number of transmembrane proteins that contain disintegrin-like and metalloproteinase-like domains. Therefore, ADAMs potentially have cell adhesion and protease activities. 14-3-3 proteins are a highly conserved family of cytoplasmic proteins that associate with several intracellular signaling molecules in the regulation of various cellular functions. Here we report the identification of a novel interaction between the ADAM 22 cytoplasmic tail and the 14-3-3ζ isoform by a yeast two-hybrid screen. The interaction between the ADAM 22 cytoplasmic tail and 14-3-3ζ was confirmed by an in vitro protein pull-down assay as well as by co-immunoprecipitation, and the binding sites were mapped to the 28 amino acid residues of the C-terminus of the ADAM 22 cytoplasmic tail. Furthermore, we found that overexpression of the ADAM 22 cytoplasmic tail in human SGH44 cells inhibited cell adhesion and spreading and that deletion or mutation of the binding site for 14-3-3ζ within the ADAM 22 cytoplasmic tail abolished the ability of the overexpressed cytoplasmic tail to alter cell adhesion and spreading. Taken together, these results for the first time demonstrate an association between ADAM 22 and a 14-3-3 protein and suggest a potential role for the 14-3-3ζ/ADAM 22 association in the regulation of cell adhesion and related signaling events. [Copyright &y& Elsevier]

Lambda-crystallin is a composition of lens in rabbit and hare. It contains the putative NAD- or FAD-binding domain, which is named as HCDH domain in 3-hydroxyacyl-CoA dehydrogenase. In our attempt to search for genes differentially expressed between liver cancer tissues and normal tissues, human CRYL1 (crystallin, lambda 1) was identified. It was downregulated in 58% of 60 Chinese HCC tissue samples. The putative protein encoded by CRYL1 shares 83% identity with rabbit lambda-crystallin and contains two HCDH domains. Interestingly, CRYL1 mRNA level is remarkably high in liver and kidney, while it is extremely low in peripheral blood leukocyte and thymus. The CRYL1mRNA levels in liver and kidney are about 1.6 and 1.2 times the total amount of that in other 14 tissues, respectively. Both the special expression pattern and the putative HCDH structure of CRYL1 suggested that the protein may be of the similar function of 3-hydroxyacyl-CoA dehydrogenase. To further understand the lambda-crystallin protein family, we cloned four novel mammalian homologs from mouse, rat, bovine and pig. The unrooted phylogenetic tree of this protein family including human and other 26 species was drawn to analyse their evolutionary relationship. In addition, human CRYL1 was mapped to chromosome 13q12.11 and mouse Cryl1 to chromosome 14 between marker D14Mit83 and D14Mit260. [Copyright &y& Elsevier]

Focuses on Wlbachia, a maternally inherited intracellular bacterium, which can infect arthropods. Association with a risk for cytoplasmic incompatibility (CI), parthenogenesis, the feminization of genetic males, and male-killing in different hosts; Investigation into the molecular mechanisms by which this bacterium causes reproductive abnormalities; Cloning and characterization of the gene encoding phage-related tail protein (PrTP) from Wolbachia in Drosophila melanogaster Cantons (wMelCS) and from Wolbachia in Drosophila melanogaster yw67c23 (wMel); Use of representational difference analysis (RDA) and ligation-mediated PCR (LM-PCR); Testing of the functionality of a bipartite nuclear localization signal sequence (NLS) in Drosophila S2 cells; Examination of PrTP expression in various strains of Wolbachia; Suggestion that PrTP has an indirect role in inducing CI; Assertion that the existence of prtp shows direct evidence of phage-mediated horizontal gene transfer (HGT) that might be linked to a variety of reproductive abnormalities of Wolbachia.

Serum and glucocorticoid-inducible kinase-like kinase (SGKL) has been identified as a new integrator that decodes lipid signals produced by the activation of phosphoinositide 3-kinase (PI3K). SGKL is activated via its lipid-binding domain (phox homology domain) in response to PI3K signaling. However, downstream targets of SGKL as well as the role of SGKL as a mediator in PI3K signaling in human tissues remain to be established. In this study, we identified human glycogen synthase kinase 3 beta (GSK-3β) as a specific interacting partner with SGKL in a yeast two-hybrid screening of human brain cDNA library. The association between these two proteins is confirmed independently in human embryonic kidney (HEK293) cells by co-immunoprecipitation. Furthermore, the kinase activity of wild-type SGKL was required for the in vitro phosphorylation of a GSK-3 crosstide fusion protein at serine-21/9 as demonstrated with a Phospho-GSK-3α/β (Ser21/9) specific antibody. The present results provide strong evidences that SGKL could utilize GSK-3β as a direct downstream target by phosphorylating GSK-3β at serine-9. [Copyright &y& Elsevier]

Protein kinase A (PKA) and neuronal nitric oxide synthase (nNOS) are important signaling molecules. It is well known that PKA can specifically phosphorylate nNOS. But the underlying molecular mechanism is still obscure. Our data indicate that the protein inhibitor of nNOS (PIN) binds to protein kinase A inhibitors (PKIs), which suggests that PKIs, together with PIN, might mediate the phosphorylation of nNOS by PKA. [Copyright &y& Elsevier]

Studies the function of the murine BRI3 gene in inducing cell death by cloning the cDNA of murine BRI3 gene from the total RNA of murine brain endothelial cells treated with tumor necrosis factor-alpha. Use of the suppression subtractive hybridization; Localization of the fusion protein in cytoplasm; Ability of the induction of the enhanced green fluorescence protein to induce cell death.

Neuronal calcium sensors (NCS) are important constituents in the intracellular signaling pathways. A novel human gene, NCS-1, was identified in the present study. Among the 16 human tissues examined, NCS-1 is expressed most abundantly in the brain. Among the brain regions, the expression level of NCS-1 in cerebral cortex is the highest, which is about six times higher than the average level of the whole brain and a hundred times higher than the spinal cord. In the 12 different anatomical regions of human brain, the expression level of NCS-1 is very high in the temporal lobe, occipital pole, frontal lobe, thalamus, amygdala and hippocampus; moderate in cerebellum, putamen, caudate nucleus; low in the medulla, substantia nigra and the lowest in corpus callosum. Our results suggest that NCS-1 in human brain might be involved in a variety of brain functions such as sensory processing, motor control, emotional control, learning and memory. [Copyright &y& Elsevier]

Studies the association of serum and glucocorticoid-inducible kinase (SGK) and 14-3-3 theta protein in serine/threonine phosphorylation mechanism for thrombopoietin/MPl signal transduction. Administration of yeast two-hybrid screening; Isolation of positive clones from independent transformants; Interaction between SGK and 14-3-3 protein.

Studies the isolation and characterization of a potential transposable element from Wolbachia, a group of Rickettsia-like bacteria. Nucleotide structure of the transposable element; Genomic difference between Wolbachia and different cytoplasmic incompatibility phenotype.

Presents a study which investigated the oxidation-reduction mechanism of methionine in proteins. Effect of peptide methionine sulfoxide reductase on the oxidation of methionine; Methodology; Results; Discussion.

Presents a study that examined the cloning of full-length circular DNA (cDNA) of a human CCT protein gene. Background on the mechanism of protein folding; Materials and methods used; Findings and implications.

Presents a study on the relationship between Reticulon (RTN) gene family and neurodoctrine lung carcinoma for cloning RTN members. Background information on lung cancer; Materials and methods used in the study; Results and discussion.

Presents a study on protein kinase B which undergoes activation in some human tumors and is involved with ras pathways. Discussion on the protein kinase present in human tissues; Materials and methods used in the study; Results and discussion.

Presents information on a study which cloned a novel gene encoding a putative human spindlin which is highly homologous to mouse spindle complementary DNA. Significance of spindlin phosphorylation; Use of Radiation Hybrid Mapping; Discussion and conclusion.

Presents a study on the cloned diseased gene of multiple neurofibromatosis type I (NF1). Methodology; Linkage analysis of NF1 using short tandem repeats (STR) as markers; Results and discussion; Conclusion.

Examines the results of a study where a single copy fragment was isolated with oligonucleotide probe from a human chromosome probe pool generated by microdissection. Short tandem repeat sequence; Polymorphic information content; Mendelian segregation.

MANSC (motif at N terminus with seven cysteines) is a novel domain with a well-conserved seven-cysteine motif that is present at the N terminus of membrane and extracellular proteins, including low-density lipoprotein receptor-related protein 11 (LRP-11), hepatocyte growth factor activator inhibitor 1 (HAI-1) and some uncharacterized proteins encoded by multicellular animals from Mollusca to Chordata. We postulate that the MANSC domain in HAI-1 might function through binding with hepatocyte growth factor activator and matriptase. [Copyright &y& Elsevier]

Discusses a study that cultured neural stem cells (NSC) isolated from the brains of embryonic mice in vitro and determined their distribution. Identification of NSC; Infection of NSC with an adenoviral vector; Expression of an exogenous reporter gene.