1887

Abstract

Mutants of resistant to mevinolin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme-A (HMGCoA) reductase (EC 1.1.1.34) were isolated and one mutant (MV71) was extensively characterized. While growth of resistant strains in the presence of mevinolin was greater than that of the wild-type strain, mevinolin at the concentration used still inhibited growth. Diploids produced by mutant/wild-type matings showed levels of mevinolin resistance which indicated incomplete dominance. Sterol synthesis in the presence of mevinolin was inhibited in strain MV71 but to a lesser degree than seen in the wild-type strain. All mevinolin resistant mutants also demonstrated a slight resistance to the antibiotic nystatin. The subcellular location of HMGCoA reductase activity in MV71 and the wild-type strain were determined and it was shown that yeast HMGCoA reductase is not regulated by a dephosphorylation mechanism as has been shown for mammalian reductases. and studies of strain MV71 and the wild-type indicated that mevinolin resistance did not result in changes in HMGCoA reductase activity as has been demonstrated in mammalian systems. Based on growth data, sterol analysis, and the lack of detection of HMGCoA reductase activity differences between strain MV71 and the wild-type, mevinolin resistance is concluded to result possibly from a mutation in , one of the two functional yeast HMGCoA reductase genes, which accounts for a minor (up to 17%) amount of total cellular reductase activity.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-134-4-1071
1988-04-01
2021-07-27
Loading full text...

Full text loading...

/deliver/fulltext/micro/134/4/mic-134-4-1071.html?itemId=/content/journal/micro/10.1099/00221287-134-4-1071&mimeType=html&fmt=ahah

References

  1. Alberts A. W., Chen J., Kuron G., Hunt V., Huff C., Hoffman C., Rothrock J., Lopez M., Joshua H., Patchett A., Monaghan R., Currie S., Stapley E., Albers-Schonberg G., Hirschfield J., Hoogsteen K., Llesch J., Springer J. 1980; Mevinolin: a highly potent competitive inhibitor of hydroxymethylglutaryl-coenzyme A reductase and a cholesterol-lowering agent. Proceedings of the National Academy of Sciences of the United States of America 77:3957–3961
    [Google Scholar]
  2. Bard M., Downing J. F. 1981; Genetic and biochemical aspects of yeast sterol regulation involving 3-hydroxy-3-methylglutaryl coenzyme A reductase. Journal of General Microbiology 125:415–420
    [Google Scholar]
  3. Basson M. E., Thorsness M., Rine J. 1986; Saccharomyces cerevisiae contains two functional genes encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase. Proceedings of the National Academy of Sciences of the United States of America 83:5563–5567
    [Google Scholar]
  4. Brown M. S., Goldstein J. L. 1980; Multivalent feedback regulation of HMGCoA reductase, a control mechanism coordinating isoprenoid synthesis and cell growth. Journal of Lipid Research 21:505–517
    [Google Scholar]
  5. Brown M. S., Faust J. R., Goldstein J. L. 1978; Induction of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in human fibroblasts incubated with compactin (ML-236B), a competitive inhibitor of the reductase. Journal of Biological Chemistry 253:1121–1128
    [Google Scholar]
  6. Chin D. J., Luskey K. L., Anderson R. G. W., Faust J. R., Goldstein J. L., Brown M. S. 1982; Appearance of crystalloid endoplasmic reticulum in compactin-resistant Chinese hamster cells with a 500-fold increase in 3-hydroxy-3- methylglutaryl coenzyme A reductase. Proceedings of the National Academy of Sciences of the United States of America 79:1185–1189
    [Google Scholar]
  7. Downing J. F., Burrows L. S., Bard M. 1980; The isolation of two mutants of Saccharomyces cerevisiae which demonstrate increased activity of 3- hydroxy-3-methylglutaryl coenzyme A reductase. Biochemical and Biophysical Research Communications 94:974–979
    [Google Scholar]
  8. Endo A., Kuroda M., Tanzawa K. 1976; Competitive inhibition of 3-hydroxy-3-methyl- glutaryl coenzyme A reductase by ML-236 and ML-236B, fungal metabolites having hypocholestero- lemic activity. FEBS Letters 72:323–326
    [Google Scholar]
  9. Edwards P. A., Lan S. F., Fogelman A. M. 1983a; Alteration in the rates of synthesis and degradation of rat liver 3-hydroxy-3-methylglutaryl coenzyme A reductase produced by cholestyramine and mevinolin. Journal of Biological Chemistry 258:10219–10222
    [Google Scholar]
  10. Edwards P. A., Lan S. F., Tanaka R., Fogelman A. M. 1983b; Mevalonalactone inhibits the rate of synthesis and enhances the rate of degradation of 3- hydroxy-3-methylglutaryl coenzyme A reductase in rat hepatocytes. Journal of Biological Chemistry 258:7272–7275
    [Google Scholar]
  11. Hardeman E. C., Jenke H. S., Simoni R. D. 1983; Overproduction of a Mr 92,000 protomer of 3- hydroxy-3-methylglutaryl-coenzyme A reductase in compactin-resistant Cl00 cells. Proceedings of the National Academy of Sciences of the United States of America 80:1516–1520
    [Google Scholar]
  12. Hardeman E. C., Endo A., Simoni R. D. 1984; Effects of compactin on the levels of 3-hydroxy-3- methylglutaryl-coenzyme A reductase in compactin- resistant Cl00 and wild type cells. Archives of Biochemistry and Biophysics 232:549–561
    [Google Scholar]
  13. Ingebritsen T. S., Gibson D. M. 1981; Assay of enzymes that modulate 3-hydroxy-3-methylglutaryl- CoA reductase by reversible phosphorylation. Methods in Enzymology 71:486–497
    [Google Scholar]
  14. Kaneko I., Hazama-Shimada Y., Endo A. 1978; Inhibitory effects on lipid metabolism in cultured cells of ML-236B, a potent inhibitor of 3-hydroxy-3- methylglutaryl coenzyme A reductase. European Journal of Biochemistry 87:313–321
    [Google Scholar]
  15. Kawaguchi A. 1970; Control of ergosterol biosynthesis in yeast. Journal of Biochemistry 67:219–227
    [Google Scholar]
  16. Kinsky S. C. 1970; Antibiotic interaction with model membranes. Annual Review of Pharmacology 10:119–142
    [Google Scholar]
  17. Lees N. D., Kemple M. D., Barbuch R. J., Smith M. A., Bard M. 1984; Differences in membrane order parameter and antibiotic sensitivity in ergo- sterol-producing strains of Saccharomyces cerevisiae. Biochimica et biophysica acta 776:105–112
    [Google Scholar]
  18. Luskey K. L., Faust J. R., Chin D. J., Brown M. S., Goldstein J. L. 1983; Amplification of the gene for 3-hydroxy-3-methylglutaryl coenzyme A reductase, but not for the 53-kDa protein in UT-1 cells. Journal of Biological Chemistry 258:8462–8469
    [Google Scholar]
  19. Molzhan S. W., Woods R. A. 1972; Polyene resistance and the isolation of sterol mutants in Saccharomyces cerevisiae. Journal of General Microbiology 72:339–348
    [Google Scholar]
  20. Nakamura C. E., Abeles R. H. 1985; Mode of interaction of 3-hydroxy-3-methylglutaryl coenzyme A reductase with strong binding inhibitors: compactin and related compounds. Biochemistry 24:1364–1376
    [Google Scholar]
  21. Parker R. A., Miller S. J., Gibson D. A. 1984; Phosphorylation of microsomal HMG-CoA reductase increases susceptibility to proteolytic degradation in vitro. Biochemical and Biophysical Research Communications 125:629–635
    [Google Scholar]
  22. Quain D. E., Haslam J. M. 1979; The effects of catabolite repression on the accumulation of steryl esters and the activity of β-hydroxymethylglutaryl- CoA reductase in Saccharomyces cerevisiae. Journal of General Microbiology 111:343–351
    [Google Scholar]
  23. Servouse M., Mons N., Baillargeat J. L., Karst F. 1984; Isolation and characterization of yeast mutants blocked in mevalonic acid formation. Biochemical and Biophysical Research Communications 123:424–430
    [Google Scholar]
  24. Sinensky M., Logel S. 1983; Inhibition of degradation of 3-hydroxy-3-methylglutaryl coenzyme A reductase by mevinolin. Journal of Biological Chemistry 258:8547–8549
    [Google Scholar]
  25. Skalnik D. G., Brown D. A., Brown P. C., Friedman R. L., Hardeman E. C., Schimke R. T., Simoni R. D. 1985; Mechanisms of 3-hydroxy-3- methylglutaryl coenzyme A reductase overaccumulation in three compactin-resistant cell lines. Journal of Biological Chemistry 260:1991–1994
    [Google Scholar]
  26. Taylor F. R., Parks L. W. 1981; An assessment of the specificity of sterol uptake and esterification in Saccharomyces cerevisiae. Journal of Biological Chemistry 256:13048–13054
    [Google Scholar]
  27. Trocha P. J., Sprinson D. B. 1976; Location and regulation of early enzymes of sterol biosynthesis in yeast. Archives of Biochemistry and Biophysics 174:45–51
    [Google Scholar]
  28. Tsujita Y., Kuroda M., Shimada Y., Tanzawa K., Arai M., Kaneko I., Tanaka M., Masuda H., Tarumi C., Watanabe Y., Fujii S. 1986; CS- 514, a competitive inhibitor of 3-hydroxy-3-methyl- glutaryl coenzyme A reductase: tissue-selective inhibitor of sterol synthesis and hypolipidemic effect on various animal species. Biochimica et biophysica acta 877:50–60
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-134-4-1071
Loading
/content/journal/micro/10.1099/00221287-134-4-1071
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error