Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Jul 19;91(15):7370–7374. doi: 10.1073/pnas.91.15.7370

Isolation and characterization of the gene encoding 2,3-oxidosqualene-lanosterol cyclase from Saccharomyces cerevisiae.

Z Shi 1, C J Buntel 1, J H Griffin 1
PMCID: PMC44401  PMID: 8041797

Abstract

The ERG7 gene encoding oxidosqualene-lanosterol cyclase [(S)-2,3-epoxysqualene mutase (cyclizing, lanosterol forming), EC 5.4.99.7] from Saccharomyces cerevisiae has been cloned by genetic complementation of a cyclase-deficient erg7 strain. The DNA sequence of this gene has been determined and found to contain an open reading frame of 2196 nt (including stop codon) that encodes a predicted protein of 731 amino acids. The predicted molecular mass of the S. cerevisiae cyclase, 83.4 kDa, is similar to the predicted molecular masses of the oxidosqualene-lanosterol cyclase from Candida albicans and the oxidosqualene-cycloartenol cyclase from Arabidopsis thaliana, as well as to the molecular masses assigned to vertebrate oxidosqualene-lanosterol cyclases; however, it is substantially larger than the molecular mass assigned to purified S. cerevisiae cyclase. At the level of DNA and predicted amino acid sequences, the S. cerevisiae and C. albicans cyclases share 56% and 63% identity, respectively. Tryptophan and tyrosine residues are unusually abundant in the predicted amino acid sequences of (oxido)-squalene cyclases, leading to a hypothesis that electron-rich aromatic side chains from these residues are essential features of cyclase active sites.

Full text

PDF
7370

Images in this article

7371Image
on p.7371

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abe I., Bai M., Xiao X. Y., Prestwich G. D. Affinity labeling of vertebrate oxidosqualene cyclases with a tritiated suicide substrate. Biochem Biophys Res Commun. 1992 Aug 31;187(1):32–38. doi: 10.1016/s0006-291x(05)81454-8. [DOI] [PubMed] [Google Scholar]
  2. Abe I., Prestwich G. D. Active site mapping of affinity-labeled rat oxidosqualene cyclase. J Biol Chem. 1994 Jan 14;269(2):802–804. [PubMed] [Google Scholar]
  3. Barton D. H., Jarman T. R., Watson K. C., Widdowson D. A., Boar R. B., Damps K. Investigations on the biosynthesis of steroids and terpenoids. Part XII. Biosynthesis of 3beta-hydroxy-triterpenoids and -steroids from (3S)-2,3-epoxy-2,3-dihydrosqualene. J Chem Soc Perkin 1. 1975;(12):1134–1138. doi: 10.1039/p19750001134. [DOI] [PubMed] [Google Scholar]
  4. Becker D. M., Guarente L. High-efficiency transformation of yeast by electroporation. Methods Enzymol. 1991;194:182–187. doi: 10.1016/0076-6879(91)94015-5. [DOI] [PubMed] [Google Scholar]
  5. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  6. Burley S. K., Petsko G. A. Amino-aromatic interactions in proteins. FEBS Lett. 1986 Jul 28;203(2):139–143. doi: 10.1016/0014-5793(86)80730-x. [DOI] [PubMed] [Google Scholar]
  7. Burley S. K., Petsko G. A. Weakly polar interactions in proteins. Adv Protein Chem. 1988;39:125–189. doi: 10.1016/s0065-3233(08)60376-9. [DOI] [PubMed] [Google Scholar]
  8. Corey E. J., Matsuda S. P., Bartel B. Isolation of an Arabidopsis thaliana gene encoding cycloartenol synthase by functional expression in a yeast mutant lacking lanosterol synthase by the use of a chromatographic screen. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11628–11632. doi: 10.1073/pnas.90.24.11628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Corey E. J., Matsuda S. P., Bartel B. Molecular cloning, characterization, and overexpression of ERG7, the Saccharomyces cerevisiae gene encoding lanosterol synthase. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2211–2215. doi: 10.1073/pnas.91.6.2211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Davies D. R., Metzger H. Structural basis of antibody function. Annu Rev Immunol. 1983;1:87–117. doi: 10.1146/annurev.iy.01.040183.000511. [DOI] [PubMed] [Google Scholar]
  11. Dougherty D. A., Stauffer D. A. Acetylcholine binding by a synthetic receptor: implications for biological recognition. Science. 1990 Dec 14;250(4987):1558–1560. doi: 10.1126/science.2274786. [DOI] [PubMed] [Google Scholar]
  12. Elledge S. J., Mulligan J. T., Ramer S. W., Spottswood M., Davis R. W. Lambda YES: a multifunctional cDNA expression vector for the isolation of genes by complementation of yeast and Escherichia coli mutations. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1731–1735. doi: 10.1073/pnas.88.5.1731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fitzgerald M., Shenk T. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs. Cell. 1981 Apr;24(1):251–260. doi: 10.1016/0092-8674(81)90521-3. [DOI] [PubMed] [Google Scholar]
  14. Gollub E. G., Liu K. P., Dayan J., Adlersberg M., Sprinson D. B. Yeast mutants deficient in heme biosynthesis and a heme mutant additionally blocked in cyclization of 2,3-oxidosqualene. J Biol Chem. 1977 May 10;252(9):2846–2854. [PubMed] [Google Scholar]
  15. Karlin S., Blaisdell B. E., Bucher P. Quantile distributions of amino acid usage in protein classes. Protein Eng. 1992 Dec;5(8):729–738. doi: 10.1093/protein/5.8.729. [DOI] [PubMed] [Google Scholar]
  16. Kelly R., Miller S. M., Lai M. H., Kirsch D. R. Cloning and characterization of the 2,3-oxidosqualene cyclase-coding gene of Candida albicans. Gene. 1990 Mar 15;87(2):177–183. doi: 10.1016/0378-1119(90)90299-7. [DOI] [PubMed] [Google Scholar]
  17. Murray V. Improved double-stranded DNA sequencing using the linear polymerase chain reaction. Nucleic Acids Res. 1989 Nov 11;17(21):8889–8889. doi: 10.1093/nar/17.21.8889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ochs D., Kaletta C., Entian K. D., Beck-Sickinger A., Poralla K. Cloning, expression, and sequencing of squalene-hopene cyclase, a key enzyme in triterpenoid metabolism. J Bacteriol. 1992 Jan;174(1):298–302. doi: 10.1128/jb.174.1.298-302.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ourisson G., Rohmer M., Poralla K. Prokaryotic hopanoids and other polyterpenoid sterol surrogates. Annu Rev Microbiol. 1987;41:301–333. doi: 10.1146/annurev.mi.41.100187.001505. [DOI] [PubMed] [Google Scholar]
  20. Pelham H. R., Hardwick K. G., Lewis M. J. Sorting of soluble ER proteins in yeast. EMBO J. 1988 Jun;7(6):1757–1762. doi: 10.1002/j.1460-2075.1988.tb03005.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  22. Ramer S. W., Elledge S. J., Davis R. W. Dominant genetics using a yeast genomic library under the control of a strong inducible promoter. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11589–11593. doi: 10.1073/pnas.89.23.11589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rohmer M., Bouvier P., Ourisson G. Molecular evolution of biomembranes: structural equivalents and phylogenetic precursors of sterols. Proc Natl Acad Sci U S A. 1979 Feb;76(2):847–851. doi: 10.1073/pnas.76.2.847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Strathern J. N., Higgins D. R. Recovery of plasmids from yeast into Escherichia coli: shuttle vectors. Methods Enzymol. 1991;194:319–329. doi: 10.1016/0076-6879(91)94024-7. [DOI] [PubMed] [Google Scholar]
  25. Strathmann M., Hamilton B. A., Mayeda C. A., Simon M. I., Meyerowitz E. M., Palazzolo M. J. Transposon-facilitated DNA sequencing. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1247–1250. doi: 10.1073/pnas.88.4.1247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sussman J. L., Harel M., Frolow F., Oefner C., Goldman A., Toker L., Silman I. Atomic structure of acetylcholinesterase from Torpedo californica: a prototypic acetylcholine-binding protein. Science. 1991 Aug 23;253(5022):872–879. doi: 10.1126/science.1678899. [DOI] [PubMed] [Google Scholar]
  27. Tam J. P. Synthetic peptide vaccine design: synthesis and properties of a high-density multiple antigenic peptide system. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5409–5413. doi: 10.1073/pnas.85.15.5409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Woolford J. L., Jr Nuclear pre-mRNA splicing in yeast. Yeast. 1989 Nov-Dec;5(6):439–457. doi: 10.1002/yea.320050604. [DOI] [PubMed] [Google Scholar]
  29. von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985 Jul 5;184(1):99–105. doi: 10.1016/0022-2836(85)90046-4. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES