Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Sep;174(17):5711–5718. doi: 10.1128/jb.174.17.5711-5718.1992

Regulation of CDP-diacylglycerol synthesis and utilization by inositol and choline in Schizosaccharomyces pombe.

P M Gaynor 1, M L Greenberg 1
PMCID: PMC206519  PMID: 1324908

Abstract

CDP-diacylglycerol (CDP-DG) is an important branchpoint intermediate in eucaryotic phospholipid biosynthesis and could be a key regulatory site in phospholipid metabolism. Therefore, we examined the effects of growth phase, phospholipid precursors, and the disruption of phosphatidylcholine (PC) synthesis on the membrane-associated phospholipid biosynthetic enzymes CDP-DG synthase, phosphatidylglycerolphosphate (PGP) synthase, phosphatidylinositol (PI) synthase, and phosphatidylserine (PS) synthase in cell extracts of the fission yeast Schizosaccharomyces pombe. In complete synthetic medium containing inositol, maximal expression of CDP-DG synthase, PGP synthase, PI synthase, and PS synthase in wild-type cells occurred in the exponential phase of growth and decreased two- to fourfold in the stationary phase of growth. In cells starved for inositol, this decrease in PGP synthase, PI synthase, and PS synthase expression was not observed. Starvation for inositol resulted in a twofold derepression of PGP synthase and PS synthase expression, while PI synthase expression decreased initially and then remained constant. Upon the addition of inositol to inositol-starved cells, there was a rapid and continued increase in PI synthase expression. We examined expression of these enzymes in cho2 and cho1 mutants, which are blocked in the methylation pathway for synthesis of PC. Choline starvation resulted in a decrease in PS synthase and CDP-DG synthase expression in cho1 but not cho2 cells. Expression of PGP synthase and PI synthase was not affected by choline starvation. Inositol starvation resulted in a 1.7-fold derepression of PGP synthase expression in cho2 but not cho1 cells when PC was synthesized. PS synthase expression was not depressed, while CDP-DG synthase and PI synthase expression decreased in cho2 and cho1 cells in the absence of inositol. These results demonstrate that (i) CDP-DG synthase, PGP synthase, PI synthase, and PS synthase are similarly regulated by growth phase; (ii) inositol affects the expression of PGP synthase, PI synthase, and PS synthase; (iii) disruption of the methylation pathway results in aberrant patterns of regulation of growth phase and phospholipid precursors. Important differences between S. pombe and Saccharomyces cerevisiae with regard to regulation of these enzymes are discussed.

Full text

PDF
5711

Selected References

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

  1. Bae-Lee M., Carman G. M. Regulation of yeast phosphatidylserine synthase and phosphatidylinositol synthase activities by phospholipids in Triton X-100/phospholipid mixed micelles. J Biol Chem. 1990 May 5;265(13):7221–7226. [PubMed] [Google Scholar]
  2. Bailis A. M., Poole M. A., Carman G. M., Henry S. A. The membrane-associated enzyme phosphatidylserine synthase is regulated at the level of mRNA abundance. Mol Cell Biol. 1987 Jan;7(1):167–176. doi: 10.1128/mcb.7.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Becker G. W., Lester R. L. Changes in phospholipids of Saccharomyces cerevisiae associated with inositol-less death. J Biol Chem. 1977 Dec 10;252(23):8684–8691. [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Carman G. M., Belunis C. J. Phosphatidylglycerophosphate synthase activity in Saccharomyces cerevisiae. Can J Microbiol. 1983 Oct;29(10):1452–1457. doi: 10.1139/m83-222. [DOI] [PubMed] [Google Scholar]
  6. Carman G. M., Henry S. A. Phospholipid biosynthesis in yeast. Annu Rev Biochem. 1989;58:635–669. doi: 10.1146/annurev.bi.58.070189.003223. [DOI] [PubMed] [Google Scholar]
  7. Carman G. M., Matas J. Solubilization of microsomal-associated phosphatidylserine synthase and phosphatidylinositol synthase from Saccharomyces cerevisiae. Can J Microbiol. 1981 Nov;27(11):1140–1149. doi: 10.1139/m81-179. [DOI] [PubMed] [Google Scholar]
  8. Carman G. M., Zaniewski R. L., Cousminer J. J. CDP-diacylglycerol synthase activity in Clostridium perfringens. Appl Environ Microbiol. 1982 Jan;43(1):81–85. doi: 10.1128/aem.43.1.81-85.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Carson M. A., Atkinson K. D., Waechter C. J. Properties of particulate and solubilized phosphatidylserine synthase activity from Saccharomyces cerevisiae. Inhibitory effect of choline in the growth medium. J Biol Chem. 1982 Jul 25;257(14):8115–8121. [PubMed] [Google Scholar]
  10. Fernandez S., Homann M. J., Henry S. A., Carman G. M. Metabolism of the phospholipid precursor inositol and its relationship to growth and viability in the natural auxotroph Schizosaccharomyces pombe. J Bacteriol. 1986 Jun;166(3):779–786. doi: 10.1128/jb.166.3.779-786.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fischl A. S., Homann M. J., Poole M. A., Carman G. M. Phosphatidylinositol synthase from Saccharomyces cerevisiae. Reconstitution, characterization, and regulation of activity. J Biol Chem. 1986 Mar 5;261(7):3178–3183. [PubMed] [Google Scholar]
  12. Gaynor P. M., Gill T., Toutenhoofd S., Summers E. F., McGraw P., Homann M. J., Henry S. A., Carman G. M. Regulation of phosphatidylethanolamine methyltransferase and phospholipid methyltransferase by phospholipid precursors in Saccharomyces cerevisiae. Biochim Biophys Acta. 1991 Nov 11;1090(3):326–332. doi: 10.1016/0167-4781(91)90197-t. [DOI] [PubMed] [Google Scholar]
  13. Gaynor P. M., Hubbell S., Schmidt A. J., Lina R. A., Minskoff S. A., Greenberg M. L. Regulation of phosphatidylglycerolphosphate synthase in Saccharomyces cerevisiae by factors affecting mitochondrial development. J Bacteriol. 1991 Oct;173(19):6124–6131. doi: 10.1128/jb.173.19.6124-6131.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Greenberg M. L., Hubbell S., Lam C. Inositol regulates phosphatidylglycerolphosphate synthase expression in Saccharomyces cerevisiae. Mol Cell Biol. 1988 Nov;8(11):4773–4779. doi: 10.1128/mcb.8.11.4773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Greenberg M. L., Klig L. S., Letts V. A., Loewy B. S., Henry S. A. Yeast mutant defective in phosphatidylcholine synthesis. J Bacteriol. 1983 Feb;153(2):791–799. doi: 10.1128/jb.153.2.791-799.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hackett J. A., Brennan P. J. The isolation and biosynthesis of the ceramide-phosphoinositol of Aspergillus niger. FEBS Lett. 1977 Mar 1;74(2):259–263. doi: 10.1016/0014-5793(77)80859-4. [DOI] [PubMed] [Google Scholar]
  17. Henry S. A., Klig L. S., Loewy B. S. The genetic regulation and coordination of biosynthetic pathways in yeast: amino acid and phospholipid synthesis. Annu Rev Genet. 1984;18:207–231. doi: 10.1146/annurev.ge.18.120184.001231. [DOI] [PubMed] [Google Scholar]
  18. Homann M. J., Bailis A. M., Henry S. A., Carman G. M. Coordinate regulation of phospholipid biosynthesis by serine in Saccharomyces cerevisiae. J Bacteriol. 1987 Jul;169(7):3276–3280. doi: 10.1128/jb.169.7.3276-3280.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Homann M. J., Henry S. A., Carman G. M. Regulation of CDP-diacylglycerol synthase activity in Saccharomyces cerevisiae. J Bacteriol. 1985 Sep;163(3):1265–1266. doi: 10.1128/jb.163.3.1265-1266.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Homann M. J., Poole M. A., Gaynor P. M., Ho C. T., Carman G. M. Effect of growth phase on phospholipid biosynthesis in Saccharomyces cerevisiae. J Bacteriol. 1987 Feb;169(2):533–539. doi: 10.1128/jb.169.2.533-539.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hubbard S. C., Brody S. Glycerophospholipid variation in choline and inositol auxotrophs of Neurospora crassa. Internal compensation among zwitterionic and anionic species. J Biol Chem. 1975 Sep 25;250(18):7173–7181. [PubMed] [Google Scholar]
  22. Karkhoff-Schweizer R. R., Kelly B. L., Greenberg M. L. Phosphatidylglycerolphosphate synthase expression in Schizosaccharomyces pombe is regulated by the phospholipid precursors inositol and choline. J Bacteriol. 1991 Oct;173(19):6132–6138. doi: 10.1128/jb.173.19.6132-6138.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kelley M. J., Bailis A. M., Henry S. A., Carman G. M. Regulation of phospholipid biosynthesis in Saccharomyces cerevisiae by inositol. Inositol is an inhibitor of phosphatidylserine synthase activity. J Biol Chem. 1988 Dec 5;263(34):18078–18085. [PubMed] [Google Scholar]
  24. Klig L. S., Homann M. J., Carman G. M., Henry S. A. Coordinate regulation of phospholipid biosynthesis in Saccharomyces cerevisiae: pleiotropically constitutive opi1 mutant. J Bacteriol. 1985 Jun;162(3):1135–1141. doi: 10.1128/jb.162.3.1135-1141.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Klig L. S., Homann M. J., Kohlwein S. D., Kelley M. J., Henry S. A., Carman G. M. Saccharomyces cerevisiae mutant with a partial defect in the synthesis of CDP-diacylglycerol and altered regulation of phospholipid biosynthesis. J Bacteriol. 1988 Apr;170(4):1878–1886. doi: 10.1128/jb.170.4.1878-1886.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lester R. L., Steiner M. R. The occurrence of diphosphoinositide and triphosphoinositide in Saccharomyces cerevisiae. J Biol Chem. 1968 Sep 25;243(18):4889–4893. [PubMed] [Google Scholar]
  27. McGraw P., Henry S. A. Mutations in the Saccharomyces cerevisiae opi3 gene: effects on phospholipid methylation, growth and cross-pathway regulation of inositol synthesis. Genetics. 1989 Jun;122(2):317–330. doi: 10.1093/genetics/122.2.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nikoloff D. M., Henry S. A. Genetic analysis of yeast phospholipid biosynthesis. Annu Rev Genet. 1991;25:559–583. doi: 10.1146/annurev.ge.25.120191.003015. [DOI] [PubMed] [Google Scholar]
  29. Summers E. F., Letts V. A., McGraw P., Henry S. A. Saccharomyces cerevisiae cho2 mutants are deficient in phospholipid methylation and cross-pathway regulation of inositol synthesis. Genetics. 1988 Dec;120(4):909–922. doi: 10.1093/genetics/120.4.909. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES