Isolation and characterization of the gene for mouse hormone-sensitive lipase.
Hormone-sensitive lipase (HSL) is the rate-limiting enzyme in hydrolysis of triglycerides in adipose tissue and of cholesteryl esters in steroidogenic tissues and macrophages. The gene encoding mouse HSL has been isolated and characterized from two overlapping lambda clones. The gene spans approximately 10.4 kb and comprises 9 exons interrupted by 8 introns. The deduced amino acid sequence specifies a protein of 759 amino acids with a Mr of 83,297 in the absence of posttranslational modifications. The known functional domains of the HSL protein are encoded by discrete exons, with the putative catalytic site (Ser423) encoded by exon 6, and the basal and regulatory phosphorylation sites (Ser557 and Ser559) encoded by exon 8. In addition, a putative lipid binding domain occurs in exon 9. The mouse protein shows 94% identity with the previously determined rat sequence and 85% identity with the recently determined human sequence. Interestingly, despite the high degree of similarity, the three species diverge significantly for a stretch of 16 amino acid residues upstream of the phosphorylation sites. In addition, an error was discovered in the carboxyl-terminal portion of the previously reported rat sequence, which produced a frame shift and premature termination of the coding sequence. The corrected rat sequence alters the identity of 12 amino acid residues and extends the protein an additional 11 residues. We have also examined the mouse HSL gene and 5' flanking region for nucleotide sequences that may modulate HSL gene transcription. Using primer extension, we identified a major transcription initiation site 593 nucleotides upstream of the protein coding sequence.(ABSTRACT TRUNCATED AT 250 WORDS)
Li
Z
Z
West Los Angeles VA Medical Center, California 90073.
Sumida
M
M
Birchbauer
A
A
Schotz
M C
MC
Reue
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary; D005091:Exons; D007438:Introns; D051379:Mice; D008969:Molecular Sequence Data; D002787:Sterol Esterase / Q000235:genetics*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D002787:Sterol Esterase