Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1996 Apr 1;24(7):1322–1329. doi: 10.1093/nar/24.7.1322

The yeast UME6 gene is required for both negative and positive transcriptional regulation of phospholipid biosynthetic gene expression.

J C Jackson 1, J M Lopes 1
PMCID: PMC145798  PMID: 8614637

Abstract

In Saccharomyces cerevisiae, regulation of the phospholipid biosynthetic genes, INO1, CHO1, CHO2 and OPI3, is known to occur at the level of transcript abundance. Derepression in response to inositol deprivation requires the INO2 and INO4 regulatory genes. Repression in response to inositol supplementation requires the OPI1 regulatory gene. Here, we examined the role of the UME6 global negative regulatory gene in expression of the phospholipid biosynthetic genes. These studies were stimulated by the finding that the INO1 promoter included a UME6 cognate cis-acting regulatory sequence (URS1). We found that the UME6 negative regulatory gene was involved in regulation of phospholipid biosynthetic gene expression through two distinct regulatory pathways. One pathway was the direct repression of INO1 expression through the URS1 element. Surprisingly, the UME6 gene was also required for derepression of CHO1, CHO2 and OPI3 gene expression. Consistent with this observation, the UME6 gene was required for wild-type levels of expression of the INO2 positive regulatory gene. Therefore, the UME6 gene has both a negative and a positive role in regulating phospholipid biosynthesis.

Full Text

The Full Text of this article is available as a PDF (162.7 KB).

Selected References

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

  1. Ambroziak J., Henry S. A. INO2 and INO4 gene products, positive regulators of phospholipid biosynthesis in Saccharomyces cerevisiae, form a complex that binds to the INO1 promoter. J Biol Chem. 1994 May 27;269(21):15344–15349. [PubMed] [Google Scholar]
  2. Ashburner B. P., Lopes J. M. Autoregulated expression of the yeast INO2 and INO4 helix-loop-helix activator genes effects cooperative regulation on their target genes. Mol Cell Biol. 1995 Mar;15(3):1709–1715. doi: 10.1128/mcb.15.3.1709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ashburner B. P., Lopes J. M. Regulation of yeast phospholipid biosynthetic gene expression in response to inositol involves two superimposed mechanisms. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9722–9726. doi: 10.1073/pnas.92.21.9722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Bowdish K. S., Mitchell A. P. Bipartite structure of an early meiotic upstream activation sequence from Saccharomyces cerevisiae. Mol Cell Biol. 1993 Apr;13(4):2172–2181. doi: 10.1128/mcb.13.4.2172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bowdish K. S., Yuan H. E., Mitchell A. P. Positive control of yeast meiotic genes by the negative regulator UME6. Mol Cell Biol. 1995 Jun;15(6):2955–2961. doi: 10.1128/mcb.15.6.2955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Buckingham L. E., Wang H. T., Elder R. T., McCarroll R. M., Slater M. R., Esposito R. E. Nucleotide sequence and promoter analysis of SPO13, a meiosis-specific gene of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9406–9410. doi: 10.1073/pnas.87.23.9406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chen D. C., Yang B. C., Kuo T. T. One-step transformation of yeast in stationary phase. Curr Genet. 1992 Jan;21(1):83–84. doi: 10.1007/BF00318659. [DOI] [PubMed] [Google Scholar]
  9. Culbertson M. R., Henry S. A. Inositol-requiring mutants of Saccharomyces cerevisiae. Genetics. 1975 May;80(1):23–40. doi: 10.1093/genetics/80.1.23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Donahue T. F., Henry S. A. Inositol Mutants of SACCHAROMYCES CEREVISIAE: Mapping the ino1 Locus and Characterizing Alleles of the ino1, ino2 and ino4 Loci. Genetics. 1981 Jul;98(3):491–503. doi: 10.1093/genetics/98.3.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Elion E. A., Warner J. R. The major promoter element of rRNA transcription in yeast lies 2 kb upstream. Cell. 1984 Dec;39(3 Pt 2):663–673. doi: 10.1016/0092-8674(84)90473-2. [DOI] [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. Greenberg M. L., Goldwasser P., Henry S. A. Characterization of a yeast regulatory mutant constitutive for synthesis of inositol-1-phosphate synthase. Mol Gen Genet. 1982;186(2):157–163. doi: 10.1007/BF00331845. [DOI] [PubMed] [Google Scholar]
  14. Guarente L., Bermingham-McDonogh O. Conservation and evolution of transcriptional mechanisms in eukaryotes. Trends Genet. 1992 Jan;8(1):27–32. doi: 10.1016/0168-9525(92)90021-u. [DOI] [PubMed] [Google Scholar]
  15. Higuchi R., Krummel B., Saiki R. K. A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res. 1988 Aug 11;16(15):7351–7367. doi: 10.1093/nar/16.15.7351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hirsch J. P., Henry S. A. Expression of the Saccharomyces cerevisiae inositol-1-phosphate synthase (INO1) gene is regulated by factors that affect phospholipid synthesis. Mol Cell Biol. 1986 Oct;6(10):3320–3328. doi: 10.1128/mcb.6.10.3320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hoshizaki D. K., Hill J. E., Henry S. A. The Saccharomyces cerevisiae INO4 gene encodes a small, highly basic protein required for derepression of phospholipid biosynthetic enzymes. J Biol Chem. 1990 Mar 15;265(8):4736–4745. [PubMed] [Google Scholar]
  18. Hudak K. A., Lopes J. M., Henry S. A. A pleiotropic phospholipid biosynthetic regulatory mutation in Saccharomyces cerevisiae is allelic to sin3 (sdi1, ume4, rpd1). Genetics. 1994 Feb;136(2):475–483. doi: 10.1093/genetics/136.2.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Johnson A. D. The price of repression. Cell. 1995 Jun 2;81(5):655–658. doi: 10.1016/0092-8674(95)90524-3. [DOI] [PubMed] [Google Scholar]
  20. Kodaki T., Hosaka K., Nikawa J., Yamashita S. Identification of the upstream activation sequences responsible for the expression and regulation of the PEM1 and PEM2 genes encoding the enzymes of the phosphatidylethanolamine methylation pathway in Saccharomyces cerevisiae. J Biochem. 1991 Feb;109(2):276–287. [PubMed] [Google Scholar]
  21. Lopes J. M., Henry S. A. Interaction of trans and cis regulatory elements in the INO1 promoter of Saccharomyces cerevisiae. Nucleic Acids Res. 1991 Jul 25;19(14):3987–3994. doi: 10.1093/nar/19.14.3987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lopes J. M., Hirsch J. P., Chorgo P. A., Schulze K. L., Henry S. A. Analysis of sequences in the INO1 promoter that are involved in its regulation by phospholipid precursors. Nucleic Acids Res. 1991 Apr 11;19(7):1687–1693. doi: 10.1093/nar/19.7.1687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lopes J. M., Schulze K. L., Yates J. W., Hirsch J. P., Henry S. A. The INO1 promoter of Saccharomyces cerevisiae includes an upstream repressor sequence (URS1) common to a diverse set of yeast genes. J Bacteriol. 1993 Jul;175(13):4235–4238. doi: 10.1128/jb.175.13.4235-4238.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Luche R. M., Smart W. C., Marion T., Tillman M., Sumrada R. A., Cooper T. G. Saccharomyces cerevisiae BUF protein binds to sequences participating in DNA replication in addition to those mediating transcriptional repression (URS1) and activation. Mol Cell Biol. 1993 Sep;13(9):5749–5761. doi: 10.1128/mcb.13.9.5749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. McGee T. P., Skinner H. B., Bankaitis V. A. Functional redundancy of CDP-ethanolamine and CDP-choline pathway enzymes in phospholipid biosynthesis: ethanolamine-dependent effects on steady-state membrane phospholipid composition in Saccharomyces cerevisiae. J Bacteriol. 1994 Nov;176(22):6861–6868. doi: 10.1128/jb.176.22.6861-6868.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nikoloff D. M., Henry S. A. Functional characterization of the INO2 gene of Saccharomyces cerevisiae. A positive regulator of phospholipid biosynthesis. J Biol Chem. 1994 Mar 11;269(10):7402–7411. [PubMed] [Google Scholar]
  27. Nikoloff D. M., McGraw P., Henry S. A. The INO2 gene of Saccharomyces cerevisiae encodes a helix-loop-helix protein that is required for activation of phospholipid synthesis. Nucleic Acids Res. 1992 Jun 25;20(12):3253–3253. doi: 10.1093/nar/20.12.3253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Park H. D., Luche R. M., Cooper T. G. The yeast UME6 gene product is required for transcriptional repression mediated by the CAR1 URS1 repressor binding site. Nucleic Acids Res. 1992 Apr 25;20(8):1909–1915. doi: 10.1093/nar/20.8.1909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Park H. O., Craig E. A. Positive and negative regulation of basal expression of a yeast HSP70 gene. Mol Cell Biol. 1989 May;9(5):2025–2033. doi: 10.1128/mcb.9.5.2025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Slekar K. H., Henry S. A. SIN3 works through two different promoter elements to regulate INO1 gene expression in yeast. Nucleic Acids Res. 1995 Jun 11;23(11):1964–1969. doi: 10.1093/nar/23.11.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Steber C. M., Esposito R. E. UME6 is a central component of a developmental regulatory switch controlling meiosis-specific gene expression. Proc Natl Acad Sci U S A. 1995 Dec 19;92(26):12490–12494. doi: 10.1073/pnas.92.26.12490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Strich R., Slater M. R., Esposito R. E. Identification of negative regulatory genes that govern the expression of early meiotic genes in yeast. Proc Natl Acad Sci U S A. 1989 Dec;86(24):10018–10022. doi: 10.1073/pnas.86.24.10018. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Strich R., Surosky R. T., Steber C., Dubois E., Messenguy F., Esposito R. E. UME6 is a key regulator of nitrogen repression and meiotic development. Genes Dev. 1994 Apr 1;8(7):796–810. doi: 10.1101/gad.8.7.796. [DOI] [PubMed] [Google Scholar]
  34. Vershon A. K., Hollingsworth N. M., Johnson A. D. Meiotic induction of the yeast HOP1 gene is controlled by positive and negative regulatory sites. Mol Cell Biol. 1992 Sep;12(9):3706–3714. doi: 10.1128/mcb.12.9.3706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Vidal M., Buckley A. M., Yohn C., Hoeppner D. J., Gaber R. F. Identification of essential nucleotides in an upstream repressing sequence of Saccharomyces cerevisiae by selection for increased expression of TRK2. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2370–2374. doi: 10.1073/pnas.92.6.2370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wang H., Clark I., Nicholson P. R., Herskowitz I., Stillman D. J. The Saccharomyces cerevisiae SIN3 gene, a negative regulator of HO, contains four paired amphipathic helix motifs. Mol Cell Biol. 1990 Nov;10(11):5927–5936. doi: 10.1128/mcb.10.11.5927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. White M. J., Hirsch J. P., Henry S. A. The OPI1 gene of Saccharomyces cerevisiae, a negative regulator of phospholipid biosynthesis, encodes a protein containing polyglutamine tracts and a leucine zipper. J Biol Chem. 1991 Jan 15;266(2):863–872. [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES