Regulation of rat liver 3-hydroxy-3-methylglutaryl coenzyme A synthase and the chromosomal localization of the human gene.
3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase was purified to homogeneity from rat liver cytoplasm. The active enzyme is a dimer composed of identical subunits of Mr = 53,000. The amino acid composition and the NH2-terminal sequence are presented. Partial cDNA clones for the enzyme were isolated by screening of a rat liver lambda gt11 expression library with antibodies raised against the purified protein. The identity of the clones was confirmed by hybrid selection and translation. When rats were fed diets supplemented with cholesterol, cholestyramine, or cholestyramine plus mevinolin, the hepatic protein mass of cytoplasmic synthase, as determined by immunoblotting, was 25, 160, and 1100%, respectively, of the mass observed in rats fed normal chow. Comparable changes in enzyme activity were observed. Approximately 9-fold increases in both HMG-CoA synthase mRNA mass and synthase mRNA activity were observed when control diets were supplemented with cholestyramine and mevinolin. When rats were fed these two drugs and then given mevalonolactone by stomach intubation, there was a 5-fold decrease of synthase mRNA within 3 h. These results indicate that cytoplasmic synthase regulation occurs primarily at the level of mRNA. This regulation is rapid and coordinate with that observed for HMG-CoA reductase. The chromosomal localization of human HMG-CoA synthase was determined by examining a panel of human-mouse somatic cell hybrids with the rat cDNA probe. Interestingly, the synthase gene resides on human chromosome 5, which has previously been shown to contain the gene for HMG-CoA reductase. Regional mapping, performed by examination of a series of chromosome 5 deletion mutants and by in situ hybridization to human chromosomes indicates that the two genes are not tightly clustered.
D000595:Amino Acid Sequence; D000818:Animals; D002792:Cholestyramine Resin / Q000494:pharmacology; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D005260:Female; D006801:Humans; D006904:Hydroxymethylglutaryl-CoA Synthase / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D008099:Liver / Q000201:enzymology*; D008148:Lovastatin; D009281:Naphthalenes / Q000494:pharmacology; D007652:Oxo-Acid-Lyases / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D014158:Transcription, Genetic
D009281:Naphthalenes; D012333:RNA, Messenger; D002792:Cholestyramine Resin; D008148:Lovastatin; D006904:Hydroxymethylglutaryl-CoA Synthase; D007652:Oxo-Acid-Lyases