title: The cDNA encoding canine dihydrolipoamide dehydrogenase contains multiple termination signals.
abstract: A 2288-bp cDNA sequence encoding dihydrolipoamide dehydrogenase (DLDH; dihydrolipoamide: NAD+ oxido-reductase; EC 1.8.1.4) was obtained by isolating a 1762-bp cDNA clone from a canine skeletal muscle library in the vector, lambda UNIZAP, combined with PCR amplification of the 5' end of the mRNA. The DLDH cDNA sequence contains a 49-bp G+C-rich 5'-untranslated region (UTR), followed by 1527 bp of coding region, and 695 bp of 3'-UTR preceding a 17-bp poly(A) tail. The single open reading frame encodes a precursor DLDH of 509 amino acids (aa) that begins with a 35-aa leader sequence. The 3'-UTR includes six possible polyadenylation signals (three AATAAA, one TATAAA and two AATGAA) and one potential stem-loop region extending from bp 1969-1991. Alignment studies of the canine and human DLDH demonstrate homology within the coding region of 98% at the aa level and 94% at the nt level. Northern blot analysis using the cDNA clone as probe showed wide tissue distribution of the mRNA, with differences in the level of expression among tissues and possible utilization of different polyadenylation sites.
authors.affiliation: Department of Brain and Vascular Research, Cleveland Clinic Foundation, OH 44195, USA.
authors.forename: A S
authors.initials: AS
authors.lastname: Martins
authors.forename: L J
authors.initials: LJ
authors.lastname: Greene
authors.forename: L L
authors.initials: LL
authors.lastname: Yoho
authors.forename: A
authors.initials: A
authors.lastname: Milsted
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D008058:Dihydrolipoamide Dehydrogenase / Q000235:genetics*; D004285:Dogs; D004792:Enzyme Precursors / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D004792:Enzyme Precursors; D012333:RNA, Messenger; D008058:Dihydrolipoamide Dehydrogenase