Search tips
Search criteria 


Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
Mol Cell Biol. 1987 September; 7(9): 3138–3146.
PMCID: PMC367948

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.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.9M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Barnard GF, Langton B, Popják G. Pseudo-isoenzyme forms of liver prenyl transferase. Biochem Biophys Res Commun. 1978 Dec 14;85(3):1097–1103. [PubMed]
  • Barnard GF, Popják G. Characterization of liver prenyl transferase and its inactivation by phenylglyoxal. Biochim Biophys Acta. 1980 Feb 22;617(2):169–182. [PubMed]
  • Barnard GF, Popják G. Human liver prenyltransferase and its characterization. Biochim Biophys Acta. 1981 Sep 15;661(1):87–99. [PubMed]
  • Benton WD, Davis RW. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. [PubMed]
  • Bergstrom JD, Wong GA, Edwards PA, Edmond J. The regulation of acetoacetyl-CoA synthetase activity by modulators of cholesterol synthesis in vivo and the utilization of acetoacetate for cholesterogenesis. J Biol Chem. 1984 Dec 10;259(23):14548–14553. [PubMed]
  • Blin N, Stafford DW. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. [PMC free article] [PubMed]
  • Brems DN, Bruenger E, Rilling HC. Isolation and characterization of a photoaffinity-labeled peptide from the catalytic site of prenyltransferase. Biochemistry. 1981 Jun 23;20(13):3711–3718. [PubMed]
  • Brems DN, Rilling HC. Photoaffinity labeling of the catalytic site of prenyltransferase. Biochemistry. 1979 Mar 6;18(5):860–864. [PubMed]
  • Brown MS, Goldstein JL. Multivalent feedback regulation of HMG CoA reductase, a control mechanism coordinating isoprenoid synthesis and cell growth. J Lipid Res. 1980 Jul;21(5):505–517. [PubMed]
  • Chan YL, Gutell R, Noller HF, Wool IG. The nucleotide sequence of a rat 18 S ribosomal ribonucleic acid gene and a proposal for the secondary structure of 18 S ribosomal ribonucleic acid. J Biol Chem. 1984 Jan 10;259(1):224–230. [PubMed]
  • Chang TY, Chang CC. Revertants of a Chinese hamster ovary cell mutant resistant to suppression by an analogue of cholesterol: isolation and partial biochemical characterization. Biochemistry. 1982 Oct 12;21(21):5316–5323. [PubMed]
  • Chang TY, Limanek JS. Regulation of cytosolic acetoacetyl coenzyme A thiolase, 3-hydroxy-3-methylglutaryl coenzyme A synthase, 3-hydroxy-3-methylglutaryl coenzyme A reductase, and mevalonate kinase by low density lipoprotein and by 25-hydroxycholesterol in Chinese hamster ovary cells. J Biol Chem. 1980 Aug 25;255(16):7787–7795. [PubMed]
  • Chin DJ, Luskey KL, Faust JR, MacDonald RJ, Brown MS, Goldstein JL. Molecular cloning of 3-hydroxy-3-methylglutaryl coenzyme a reductase and evidence for regulation of its mRNA. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7704–7708. [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Church GM, Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. [PubMed]
  • Clarke CF, Edwards PA, Lan SF, Tanaka RD, Fogelman AM. Regulation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase mRNA levels in rat liver. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3305–3308. [PubMed]
  • Clarke CF, Fogelman AM, Edwards PA. Diurnal rhythm of rat liver mRNAs encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase. Correlation of functional and total mRNA levels with enzyme activity and protein. J Biol Chem. 1984 Aug 25;259(16):10439–10447. [PubMed]
  • Clarke CF, Fogelman AM, Edwards PA. Transcriptional regulation of the 3-hydroxy-3-methylglutaryl coenzyme A reductase gene in rat liver. J Biol Chem. 1985 Nov 15;260(26):14363–14367. [PubMed]
  • Cochran BH, Reffel AC, Stiles CD. Molecular cloning of gene sequences regulated by platelet-derived growth factor. Cell. 1983 Jul;33(3):939–947. [PubMed]
  • Collins FS, Weissman SM. The molecular genetics of human hemoglobin. Prog Nucleic Acid Res Mol Biol. 1984;31:315–462. [PubMed]
  • Deininger PL. Approaches to rapid DNA sequence analysis. Anal Biochem. 1983 Dec;135(2):247–263. [PubMed]
  • Dihanich ME, Najarian D, Clark R, Gillman EC, Martin NC, Hopper AK. Isolation and characterization of MOD5, a gene required for isopentenylation of cytoplasmic and mitochondrial tRNAs of Saccharomyces cerevisiae. Mol Cell Biol. 1987 Jan;7(1):177–184. [PMC free article] [PubMed]
  • Edwards PA, Lan SF, Fogelman AM. Alterations in the rates of synthesis and degradation of rat liver 3-hydroxy-3-methylglutaryl coenzyme A reductase produced by cholestyramine and mevinolin. J Biol Chem. 1983 Sep 10;258(17):10219–10222. [PubMed]
  • Edwards PA, Lan SF, Tanaka RD, Fogelman AM. Mevalonolactone inhibits the rate of synthesis and enhances the rate of degradation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in rat hepatocytes. J Biol Chem. 1983 Jun 25;258(12):7272–7275. [PubMed]
  • Faust JR, Brown MS, Goldstein JL. Synthesis of delta 2-isopentenyl tRNA from mevalonate in cultured human fibroblasts. J Biol Chem. 1980 Jul 25;255(14):6546–6548. [PubMed]
  • Faust JR, Goldstein JL, Brown MS. Synthesis of ubiquinone and cholesterol in human fibroblasts: regulation of a branched pathway. Arch Biochem Biophys. 1979 Jan;192(1):86–99. [PubMed]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed]
  • Gil G, Goldstein JL, Slaughter CA, Brown MS. Cytoplasmic 3-hydroxy-3-methylglutaryl coenzyme A synthase from the hamster. I. Isolation and sequencing of a full-length cDNA. J Biol Chem. 1986 Mar 15;261(8):3710–3716. [PubMed]
  • Gil G, Brown MS, Goldstein JL. Cytoplasmic 3-hydroxy-3-methylglutaryl coenzyme A synthase from the hamster. II. Isolation of the gene and characterization of the 5' flanking region. J Biol Chem. 1986 Mar 15;261(8):3717–3724. [PubMed]
  • Gould RG, Swyryd EA. Sites of control of hepatic cholesterol biosynthesis. J Lipid Res. 1966 Sep;7(5):698–707. [PubMed]
  • Gubler U, Hoffman BJ. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. [PubMed]
  • Hall RH. N6-(delta 2-isopentenyl)adenosine: chemical reactions, biosynthesis, metabolism, and significance to the structure and function of tRNA. Prog Nucleic Acid Res Mol Biol. 1970;10:57–86. [PubMed]
  • Itakura K, Rossi JJ, Wallace RB. Synthesis and use of synthetic oligonucleotides. Annu Rev Biochem. 1984;53:323–356. [PubMed]
  • Kersten H. On the biological significance of modified nucleosides in tRNA. Prog Nucleic Acid Res Mol Biol. 1984;31:59–114. [PubMed]
  • Lee KL, Isham KR, Stringfellow L, Rothrock R, Kenney FT. Molecular cloning of cDNAs cognate to genes sensitive to hormonal control in rat liver. J Biol Chem. 1985 Dec 25;260(30):16433–16438. [PubMed]
  • Lehrach H, Diamond D, Wozney JM, Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. [PubMed]
  • Liscum L, Luskey KL, Chin DJ, Ho YK, Goldstein JL, Brown MS. Regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase and its mRNA in rat liver as studied with a monoclonal antibody and a cDNA probe. J Biol Chem. 1983 Jul 10;258(13):8450–8455. [PubMed]
  • Luskey KL, Chin DJ, MacDonald RJ, Liscum L, Goldstein JL, Brown MS. Identification of a cholesterol-regulated 53,000-dalton cytosolic protein in UT-1 cells and cloning of its cDNA. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6210–6214. [PubMed]
  • Maxson R, Cohn R, Kedes L, Mohun T. Expression and organization of histone genes. Annu Rev Genet. 1983;17:239–277. [PubMed]
  • McClanahan T, McEntee K. Specific transcripts are elevated in Saccharomyces cerevisiae in response to DNA damage. Mol Cell Biol. 1984 Nov;4(11):2356–2363. [PMC free article] [PubMed]
  • Mehrabian M, Callaway KA, Clarke CF, Tanaka RD, Greenspan M, Lusis AJ, Sparkes RS, Mohandas T, Edmond J, Fogelman AM, et al. Regulation of rat liver 3-hydroxy-3-methylglutaryl coenzyme A synthase and the chromosomal localization of the human gene. J Biol Chem. 1986 Dec 5;261(34):16249–16255. [PubMed]
  • Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. [PubMed]
  • Osborne TF, Goldstein JL, Brown MS. 5' end of HMG CoA reductase gene contains sequences responsible for cholesterol-mediated inhibition of transcription. Cell. 1985 Aug;42(1):203–212. [PubMed]
  • Parnes JR, Velan B, Felsenfeld A, Ramanathan L, Ferrini U, Appella E, Seidman JG. Mouse beta 2-microglobulin cDNA clones: a screening procedure for cDNA clones corresponding to rare mRNAs. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2253–2257. [PubMed]
  • Pelletier J, Sonenberg N. Insertion mutagenesis to increase secondary structure within the 5' noncoding region of a eukaryotic mRNA reduces translational efficiency. Cell. 1985 Mar;40(3):515–526. [PubMed]
  • Reed BC, Rilling HC. Substrate Binding of avian liver prenyltransferase. Biochemistry. 1976 Aug 24;15(17):3739–3745. [PubMed]
  • Reynolds GA, Basu SK, Osborne TF, Chin DJ, Gil G, Brown MS, Goldstein JL, Luskey KL. HMG CoA reductase: a negatively regulated gene with unusual promoter and 5' untranslated regions. Cell. 1984 Aug;38(1):275–285. [PubMed]
  • Reynolds GA, Goldstein JL, Brown MS. Multiple mRNAs for 3-hydroxy-3-methylglutaryl coenzyme A reductase determined by multiple transcription initiation sites and intron splicing sites in the 5'-untranslated region. J Biol Chem. 1985 Aug 25;260(18):10369–10377. [PubMed]
  • Rilling HC. Eukaryotic prenyltransferases. Methods Enzymol. 1985;110:145–152. [PubMed]
  • Rodwell VW, Nordstrom JL, Mitschelen JJ. Regulation of HMG-CoA reductase. Adv Lipid Res. 1976;14:1–74. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Skalnik DG, Brown DA, Brown PC, Friedman RL, Hardeman EC, Schimke RT, Simoni RD. Mechanisms of 3-hydroxy-3-methylglutaryl coenzyme A reductase overaccumulation in three compactin-resistant cell lines. J Biol Chem. 1985 Feb 25;260(4):1991–1994. [PubMed]
  • Slakey LL, Craig MC, Beytia E, Briedis A, Feldbruegge DH, Dugan RE, Qureshi AA, Subbarayan C, Porter JW. The effects of fasting, refeeding, and time of day on the levels of enzymes effecting the conversion of -hydroxy- -methylglutaryl-coenzyme A to squalene. J Biol Chem. 1972 May 25;247(10):3014–3022. [PubMed]
  • St John TP, Davis RW. Isolation of galactose-inducible DNA sequences from Saccharomyces cerevisiae by differential plaque filter hybridization. Cell. 1979 Feb;16(2):443–452. [PubMed]
  • Weinberg RA, Penman S. Processing of 45 s nucleolar RNA. J Mol Biol. 1970 Jan 28;47(2):169–178. [PubMed]
  • White LW, Rudney H. Regulation of 3-hydroxy-3-methylglutarate and mevalonate biosynthesis by rat liver homogenates. Effects of fasting, cholesterol feeding, and triton administration. Biochemistry. 1970 Jun 23;9(13):2725–2731. [PubMed]
  • Young RA, Davis RW. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. [PubMed]
  • Young RA, Davis RW. Yeast RNA polymerase II genes: isolation with antibody probes. Science. 1983 Nov 18;222(4625):778–782. [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)