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The chaperonin containing t-complex polypeptide 1 (TCP-1), as one of its subunits, CCT, is a cytosolic heterooligomeric molecular chaperone assisting in the folding of proteins in eukaryotic cytosol. We have isolated a Tcp-1-related 119-bp cDNA fragment from a human cDNA library by polymerase chain reaction, and cloned full-length mouse cDNAs orthologous to the human cDNA by hybridization. The nucleotide...
A reporter plasmid coding for the low-molecular-weight (low-M r ) form of single-chain urokinase-type plasminogen activator (low-M r u-PA; Leu 144 -Leu 411 ) has been constructed and used to analyze promoter activity. Vectors containing the low-M r u-PA cDNA coupled to hormone-responsive promoters were introduced into mammalian cells. Following...
Exon trapping allows for the rapid identification and cloning of coding regions from cloned eukaryotic DNA. In preliminary experiments, we observed two phenomena which limited the exon-trapping efficiency of pSPL3-based systems. The first factor that affected performance was revealed when we found that up to 50% of the putative trapped exons contained sequences derived from the intron of the pSPL3...
We have isolated a cDNA corresponding to the hamster peroxisome proliferator-activated receptor haPPARγ, a member of the steroid nuclear hormone receptor superfamily of transcription factors. haPPARγ mRNA is highly expressed in adipose tissue, and is expressed in lung, heart, kidney, liver and spleen to a lower extent. Thus, haPPARγ may function in activating the transcription of target genes in a...
Interleukin-3 (IL-3) is one of the cytokines that act during the early and late stages of blood cell formation. To enable the study of the role of IL-3 in bovine haemopoietic stem cell differentiation, the polymerase chain reaction was used to amplify an IL-3 cDNA from first-strand cDNAs prepared from RNA isolated from 4- and 5-hour concanavalin-A-stimulated peripheral blood lymphocytes (PBL) from...
The complete coding sequence of a mouse glutamate transporter (mEAAT2) has been cloned by polymerase chain reaction (PCR) from adult whole-brain total RNA. Southern hybridization analysis of PCR products amplified from templates derived from various murine adult tissues demonstrated that the trancript for mEAAT2 was specific to the central nervous system. High-affinity transport of d-aspartate, K...
In the pond snail Lymnaea stagnalis (Ls), growth and associated processes are likely to be controlled by a family of molluscan insulin-related peptides (MIP). Here we report on the cloning of a cDNA encoding a putative receptor for these MIP. This cDNA was isolated from Ls via PCR with degenerate oligodeoxynucleotides corresponding to conserved parts of the tyrosine kinase domain of the human insulin...
Members of the recently inserted human-specific (HS)/predicted variant (PV) subfamily of Alu elements were sequenced. A number of these Alu elements share greater than 98% sequence identity with the subfamily consensus sequence, and they are flanked by perfect 5 and 3 direct repeats ranging in size from 6 to 15 nucleotides (nt). Based on the low number of random mutations, the estimated average...
Transcripts of the gene encoding the p65 subunit of the NF-κB/Rel transcription factor complex have been reported to undergo alternative splicing to generate one derivative lacking codons for amino acids (aa) 222 to 231 (p65Δ1) and another that lacks codons for aa 13 to 25 (p65Δ2) of the conserved Rel homology domain [Narayaran et al., Science 256 (1992) 317-320; Lyle et al., Gene 138 (1994) 265-266]...
The synthesis of the human topoisomerase I (Topo I) in Escherichia coli (Ec) results in phenotypes consistent with the appearance of a DNA-relaxing activity. High-level expression was problematic and recloning the gene in a reduced-copy-number plasmid under more stringently regulated control produced a stable plasmid. An Ec strain with an inducible sensitivity to the eukaryotic Topo I poison, camptothecin...
The human aldehyde dehydrogenase gene (ALDH) family in characterized by two major conserved DNA sequences encoding residues which are possibly involved in the catalytic function and the maintenance of the functional conformation of the ALDH enzyme. This property is the basis for synthesizing the degenerate primers to clone several cDNAs of the ALDH isozymes. In this report, we describe the cDNA...
The nucleotide sequence of the polyubiquitin gene UbC of HeLa S3 cells and its upstream region was determined and characterized. Recognition sequences for the transcription factors HSF, NF B, AP-1((c-jun), NF-IL6 and Sp1 were found in the upstream control region, a result consistent with the observation of a distinct regulatory response for the UbC gene compared with that of another polyubiquitin...
Mutations in the mouse quaking locus can result in two different types of developmental phenotypes: (1) a deficiency of myelin in the central nervous system that is accompanied by a characteristic tremor, or (2) embryonic lethality around day 9 of gestation. A quaking candidate gene (qkI) that encodes a KH motif protein has recently been identified. We have isolated and characterized cDNAs encoding...
The structure of two human aldehyde dehydrogenase genes, ALDH7 and ALDH8, have been determined. The ALDH7 gene spans about 20 kb of the human genomic DNA and is composed of 9 coding exons. The ALDH8 gene is over 10 kb in length and consists of at least 10 exons. The ALDH8 gene contains an in-frame stop codon at the 17th codon position from the first initiator Met. The coding region of the ALDH7 gene...
SECIS elements form stem-loop structures in the 3 untranslated regions (UTR) of eukaryotic mRNAs that encode selenoproteins. These elements direct incorporation of selenocysteine at UGA codons, provided the SECIS element lies a sufficient distance from the UGA. The cDNAs encoding skeletal muscle selenoprotein W from human, rhesus monkey, sheep, rat, and mouse contained highly similar SECIS elements...
We have isolated cDNAs of the human gene for high mobility group protein HMG2a, using the method of direct cDNA selection. The gene maps to chromosome band Xq28, and is located within 40kb from marker DXS1684, at a distance of ~5.4Mb from the telomere. The deduced human HMG2a protein sequence has a length of 199 amino acids and is 97% identical to the sequence of chicken HMG2a. The 3' untranslated...
A highly conserved laminin receptor processed pseudogene (LAMRL5) that has been isolated from a fetal brain cDNA library is described. The pseudogene is a complete copy (97.9% identical) of the transcribed laminin receptor (LAMR1) with all the introns precisely removed. The sequence has direct repeats of 18bp at either end. It has an 885 nucleotide open reading frame from the start methionine codon...
Human SLIM1 is a recently described gene of the LIM-only class encoding four and a half tandemly repeated LIM domains. LIM domains are double zinc finger structures which provide an interface for protein/protein interactions and are conserved in a variety of nuclear and cytoplasmic factors important in cell fate determination and cellular regulation. Here we report the structural organization, expression...
Nucleotide excision repair (NER) is found throughout nature, in eubacteria, eukaryotes and archaea. In human cells it is the main pathway for the removal of damage caused by UV light, but it also acts on a wide variety of other bulky helix-distorting lesions caused by chemical mutagens. An ongoing challenge is to understand how a site of DNA damage is located during NER and distinguished from non-damaged...
In an attempt to identify novel chemokine receptor genes, cDNA expressed sequence tags (EST) were analyzed for a significant homology with mammalian chemokine receptors. The sequence from one of the selected EST clones was used to generate a full-length cDNA encoding a putative seven transmembrane receptor, CCRL1-CC chemokine receptor like 1. The full-length receptor encodes a polypeptide of 350 amino...
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