Molecular cloning and sequence of a cholesterol-repressible enzyme related to prenyltransferase in the isoprene biosynthetic pathway.
Differential hybridization and molecular cloning have been used to isolate CR39, a cDNA which hybridizes to a 1.2-kilobase (kb) mRNA in rat liver. The level of CR39 mRNA was increased seven- to ninefold over normal levels by dietary cholestyramine and mevinolin and decreased about fourfold compared with normal levels by cholesterol feeding or administration of mevalonate. Similar changes in the mRNA levels of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and HMG-CoA synthase were observed under the various conditions. In vitro translation of either CR39 hybrid selected RNA or 1.2-kb CR39 RNA generated by an SP6 in vitro transcription system produced a polypeptide of 39,000 daltons. As deduced from the nucleotide sequence of a full-length CR39 cDNA, the rat CR39 polypeptide contained 344 amino acids and had a molecular weight of 39,615. The predicted amino acid composition and submit molecular weight of the rat CR39 were very similar to those of prenyltransferases isolated from chicken, pig, and human. The sequence of amino acid residues 173 through 203 in the rat CR39 polypeptide showed that 17 out of 30 matched an active-site peptide of avian liver prenyltransferase. Thus, alterations in the rate of cholesterogenesis resulted in the coordinate regulation of three mRNAs encoding HMG-CoA reductase, HMG-CoA synthase, and CR39, the latter being tentatively identified as prenyltransferase.
Clarke
C F
CF
Department of Medicine, School of Medicine, University of California, Los Angeles 90024.
Tanaka
R D
RD
Svenson
K
K
Wamsley
M
M
Fogelman
A M
AM
Edwards
P A
PA
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001483:Base Sequence; D002784:Cholesterol / Q000502:physiology*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004122:Dimethylallyltranstransferase / Q000235:genetics*; D004794:Enzyme Repression; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D014166:Transferases / Q000235:genetics*
D000596:Amino Acids; D004247:DNA; D002784:Cholesterol; D014166:Transferases; D004122:Dimethylallyltranstransferase