We isolated three human testis-specific genes by a differential mRNA display method. The cDNAs contained open reading frames of 1620, 453, and 333 nucleotides, encoding 540, 151, and 111 amino acids, respectively. The first of these genes, designated TSA303, encodes a novel protein homologous to TCP20, one of the subunits of the human TRiC chaperonin complex that can bind newly synthesized or unstable folding intermediates of polypeptides and assist substrate proteins in folding, assembly, and transport. The second, TSA806, encodes a novel protein containing 3.3 contiguous repeats of the cdc10/swi6 (ankyrin) motif that was originally found in products of cell cycle control genes of yeast and cell fate determination genes in Drosophila and Caenorhabditis elegans. The third gene, TSA903, encodes a protein homologous to the C-terminal region of murine uridine monophosphate kinase. Northern blot analysis confirmed that in 16 human adult tissues examined, each of these genes was expressed specifically in the testis. From the results of cDNA screening of nearly 1 million plaques, the abundance of each transcript in a preparation of total mRNA was estimated as 0.0004% (TSA303), 0.0006% (TSA806), and 0.0002% (TSA903). Our results imply that the differential display method is a powerful tool for isolation of tissue-specific genes even if they are expressed at a level as low as 1 in several hundred thousand to a million molecules of total mRNA.
"Instead, the mRNAs in this category are chaperones, intracellular membrane proteins, cytoskeletal elements, or carry signals in sperm. Cct6B is part of the TCP1 ring complex that folds cytoplasmic proteins such as actin, and carries proteins to targets, even down axons (Ozaki et al., 1996; Bourke et al., 2002). Calmegin (Clgn) is a chaperone that helps dimerize the sperm proteins fertilin a and fertilin h, a function thought to be critical because Clgn null males are sterile (Nakanishi et al., 2004). "
[Show abstract][Hide abstract] ABSTRACT: The olfactory epithelium has the unusual ability to replace its neurons.We forced replacement of mouse olfactory sensory neurons by bulbectomy. Microarray, bioinformatics, and in situ hybridization techniques detected a rapid shift in favor of pro-apoptotic proteins, a progressive immune response by macrophages and dendritic cells, and identified or predicted 439 mRNAs enriched in olfactory sensory neurons, including gene silencing factors and sperm flagellar proteins. Transcripts encoding cell cycle regulators, axonogenesis proteins, and transcription factors and signaling proteins that promote proliferation and differentiation were increased at 5-7 days after bulbectomy and were expressed by basal progenitor cells or immature neurons. The transcription factors included Nhlhl, Hes6, Lmycl, c-Myc, Mxd4, Idl,Nmycl, Cited2, c-Myb, Mybll, Tead2, Dpl, Gata2, Lmol, and Soxll. The data reveal significant similarities with embryonic neurogenesis and make several mechanistic predictions, including the roles of the transcription factors in the olfactory sensory neuron lineage.
"Because the polymerase chain reaction (PCR) products contained plural bands of the same size, the reamplified products were electrophoresed on agarose gels containing a bisbenzimide–PEG (polyethylene glycol) conjugate, H.A.-Yellow (Hanse Analytik GmbH, Bremen, Germany ); as this reagent specifically adheres to adenine/ thymine (A/T) bases, target genes can be separated from background bands. The desired cDNA fragments were subcloned in the manner described previously (Ozaki et al. 1996). Nucleotide sequences were determined with an ABI 377 auto-sequencer (Applied Biosystems, Foster City, CA, USA). "
[Show abstract][Hide abstract] ABSTRACT: We have isolated the complete cDNA of a human SEL-1L gene, termed TSA305, that is abundantly expressed only in the pancreas. The cDNA contained an open reading frame of 2382 nucleotides, encoding a deduced protein of 794 amino acids whose predicted sequence showed 46% identity and 64% similarity with SEL-1 of Caenorhabditis elegans. SEL-1 is thought to be a negative regulator of the NOTCH, LIN-12, and GLP-1 receptors, which are required for differentiation and maturation of cells as well as cell-cell interactions during development in C. elegans. The degree of homology among these proteins suggests that the TSA305 gene product may be a member of the SEL-1 family and therefore involved in downregulation of mammalian Notch signaling. Direct sequencing revealed at least 20 coding exons in TSA305. We localized the gene to chromosome bands 14q24.3-q31 by radiation hybrid (RH) mapping and fluorescence in situ hybridization (FISH). The IDDM11 locus has been mapped in this region, and TSA305 may represent a candidate gene for predisposition in some families whose insulin-dependent diabetes is not linked to the HLA locus.
Journal of Human Genetics 02/1999; 44(5):330-6. DOI:10.1007/s100380050171 · 2.46 Impact Factor
"Aminoglycoside-aminocyclitol antibiotics form a large family of antimicrobial agents, including streptmycin, gentamicin, kanamycin, and amikacin, which act principally against gram-negative bacteria (Davis, 1986). Plasmidencoded resistance to these agents is mediated by enzymatic modification; i. e., by acetylation of amino groups or by adenylation or phosphorylation of hydroxyl residues (Haas and Dowding 1975; Mitsuhashi, 1982). Differential display (Liang and Pardee 1992; Liang et al. 1992; Bauer et al. 1993; Utans et al. 1994; Donohue et al. 1995) is an effective method for isolating genes that are expressed differentialy among separate tissues. "
[Show abstract][Hide abstract] ABSTRACT: Using the differential display method to detect tissue-specific genes, we isolated a novel human gene, designated TSC501, that is expressed in kidney and liver. The cDNA contained an open reading frame of 681 nucleotides encoding 227 amino acids. The predicted product showed homologies in amino acid sequence to three different bacterial acetyltransferases, enzymes that are involved in drug resistance. Radiation hybrid mapping localized this gene to human chromosome bands 2p12-p13.1.
Journal of Human Genetics 02/1998; 43(4):255-8. DOI:10.1007/s100380050084 · 2.46 Impact Factor
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