Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1994 Jun;14(6):4135–4144. doi: 10.1128/mcb.14.6.4135

GPD1, which encodes glycerol-3-phosphate dehydrogenase, is essential for growth under osmotic stress in Saccharomyces cerevisiae, and its expression is regulated by the high-osmolarity glycerol response pathway.

J Albertyn 1, S Hohmann 1, J M Thevelein 1, B A Prior 1
PMCID: PMC358779  PMID: 8196651

Abstract

The yeast Saccharomyces cerevisiae responds to osmotic stress, i.e., an increase in osmolarity of the growth medium, by enhanced production and intracellular accumulation of glycerol as a compatible solute. We have cloned a gene encoding the key enzyme of glycerol synthesis, the NADH-dependent cytosolic glycerol-3-phosphate dehydrogenase, and we named it GPD1. gpd1 delta mutants produced very little glycerol, and they were sensitive to osmotic stress. Thus, glycerol production is indeed essential for the growth of yeast cells during reduced water availability. hog1 delta mutants lacking a protein kinase involved in osmostress-induced signal transduction (the high-osmolarity glycerol response [HOG] pathway) failed to increase glycerol-3-phosphate dehydrogenase activity and mRNA levels when osmotic stress was imposed. Thus, expression of GPD1 is regulated through the HOG pathway. However, there may be Hog1-independent mechanisms mediating osmostress-induced glycerol accumulation, since a hog1 delta strain could still enhance its glycerol content, although less than the wild type. hog1 delta mutants are more sensitive to osmotic stress than isogenic gpd1 delta strains, and gpd1 delta hog1 delta double mutants are even more sensitive than either single mutant. Thus, the HOG pathway most probably has additional targets in the mechanism of adaptation to hypertonic medium.

Full text

PDF
4135

Images in this article

4140Image
on p.4140
4140Image
on p.4140
4141Image
on p.4141
4141Image
on p.4141
4142Image
on p.4142

Selected References

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

  1. Adler L., Blomberg A., Nilsson A. Glycerol metabolism and osmoregulation in the salt-tolerant yeast Debaryomyces hansenii. J Bacteriol. 1985 Apr;162(1):300–306. doi: 10.1128/jb.162.1.300-306.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albertyn J., Hohmann S., Prior B. A. Characterization of the osmotic-stress response in Saccharomyces cerevisiae: osmotic stress and glucose repression regulate glycerol-3-phosphate dehydrogenase independently. Curr Genet. 1994 Jan;25(1):12–18. doi: 10.1007/BF00712960. [DOI] [PubMed] [Google Scholar]
  3. Albertyn J., van Tonder A., Prior B. A. Purification and characterization of glycerol-3-phosphate dehydrogenase of Saccharomyces cerevisiae. FEBS Lett. 1992 Aug 17;308(2):130–132. doi: 10.1016/0014-5793(92)81259-o. [DOI] [PubMed] [Google Scholar]
  4. André L., Hemming A., Adler L. Osmoregulation in Saccharomyces cerevisiae. Studies on the osmotic induction of glycerol production and glycerol-3-phosphate dehydrogenase (NAD+) FEBS Lett. 1991 Jul 29;286(1-2):13–17. doi: 10.1016/0014-5793(91)80930-2. [DOI] [PubMed] [Google Scholar]
  5. Bell W., Klaassen P., Ohnacker M., Boller T., Herweijer M., Schoppink P., Van der Zee P., Wiemken A. Characterization of the 56-kDa subunit of yeast trehalose-6-phosphate synthase and cloning of its gene reveal its identity with the product of CIF1, a regulator of carbon catabolite inactivation. Eur J Biochem. 1992 Nov 1;209(3):951–959. doi: 10.1111/j.1432-1033.1992.tb17368.x. [DOI] [PubMed] [Google Scholar]
  6. Bennetzen J. L., Hall B. D. Codon selection in yeast. J Biol Chem. 1982 Mar 25;257(6):3026–3031. [PubMed] [Google Scholar]
  7. Berben G., Dumont J., Gilliquet V., Bolle P. A., Hilger F. The YDp plasmids: a uniform set of vectors bearing versatile gene disruption cassettes for Saccharomyces cerevisiae. Yeast. 1991 Jul;7(5):475–477. doi: 10.1002/yea.320070506. [DOI] [PubMed] [Google Scholar]
  8. Blomberg A., Adler L. Physiology of osmotolerance in fungi. Adv Microb Physiol. 1992;33:145–212. doi: 10.1016/s0065-2911(08)60217-9. [DOI] [PubMed] [Google Scholar]
  9. Boguslawski G. PBS2, a yeast gene encoding a putative protein kinase, interacts with the RAS2 pathway and affects osmotic sensitivity of Saccharomyces cerevisiae. J Gen Microbiol. 1992 Nov;138(11):2425–2432. doi: 10.1099/00221287-138-11-2425. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Brewster J. L., de Valoir T., Dwyer N. D., Winter E., Gustin M. C. An osmosensing signal transduction pathway in yeast. Science. 1993 Mar 19;259(5102):1760–1763. doi: 10.1126/science.7681220. [DOI] [PubMed] [Google Scholar]
  12. Chowdhury S., Smith K. W., Gustin M. C. Osmotic stress and the yeast cytoskeleton: phenotype-specific suppression of an actin mutation. J Cell Biol. 1992 Aug;118(3):561–571. doi: 10.1083/jcb.118.3.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Christianson T. W., Sikorski R. S., Dante M., Shero J. H., Hieter P. Multifunctional yeast high-copy-number shuttle vectors. Gene. 1992 Jan 2;110(1):119–122. doi: 10.1016/0378-1119(92)90454-w. [DOI] [PubMed] [Google Scholar]
  14. Csonka L. N., Hanson A. D. Prokaryotic osmoregulation: genetics and physiology. Annu Rev Microbiol. 1991;45:569–606. doi: 10.1146/annurev.mi.45.100191.003033. [DOI] [PubMed] [Google Scholar]
  15. Dobson D. E., Groves D. L., Spiegelman B. M. Nucleotide sequence and hormonal regulation of mouse glycerophosphate dehydrogenase mRNA during adipocyte and muscle cell differentiation. J Biol Chem. 1987 Feb 5;262(4):1804–1809. [PubMed] [Google Scholar]
  16. Errede B., Levin D. E. A conserved kinase cascade for MAP kinase activation in yeast. Curr Opin Cell Biol. 1993 Apr;5(2):254–260. doi: 10.1016/0955-0674(93)90112-4. [DOI] [PubMed] [Google Scholar]
  17. Gancedo C., Gancedo J. M., Sols A. Glycerol metabolism in yeasts. Pathways of utilization and production. Eur J Biochem. 1968 Jul;5(2):165–172. doi: 10.1111/j.1432-1033.1968.tb00353.x. [DOI] [PubMed] [Google Scholar]
  18. Gaxiola R., de Larrinoa I. F., Villalba J. M., Serrano R. A novel and conserved salt-induced protein is an important determinant of salt tolerance in yeast. EMBO J. 1992 Sep;11(9):3157–3164. doi: 10.1002/j.1460-2075.1992.tb05392.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
  20. Gläser H. U., Thomas D., Gaxiola R., Montrichard F., Surdin-Kerjan Y., Serrano R. Salt tolerance and methionine biosynthesis in Saccharomyces cerevisiae involve a putative phosphatase gene. EMBO J. 1993 Aug;12(8):3105–3110. doi: 10.1002/j.1460-2075.1993.tb05979.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hohmann S., Neves M. J., de Koning W., Alijo R., Ramos J., Thevelein J. M. The growth and signalling defects of the ggs1 (fdp1/byp1) deletion mutant on glucose are suppressed by a deletion of the gene encoding hexokinase PII. Curr Genet. 1993;23(4):281–289. doi: 10.1007/BF00310888. [DOI] [PubMed] [Google Scholar]
  22. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kaasen I., Falkenberg P., Styrvold O. B., Strøm A. R. Molecular cloning and physical mapping of the otsBA genes, which encode the osmoregulatory trehalose pathway of Escherichia coli: evidence that transcription is activated by katF (AppR) J Bacteriol. 1992 Feb;174(3):889–898. doi: 10.1128/jb.174.3.889-898.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kendall R. L., Yamada R., Bradshaw R. A. Cotranslational amino-terminal processing. Methods Enzymol. 1990;185:398–407. doi: 10.1016/0076-6879(90)85035-m. [DOI] [PubMed] [Google Scholar]
  25. Larsson K., Ansell R., Eriksson P., Adler L. A gene encoding sn-glycerol 3-phosphate dehydrogenase (NAD+) complements an osmosensitive mutant of Saccharomyces cerevisiae. Mol Microbiol. 1993 Dec;10(5):1101–1111. doi: 10.1111/j.1365-2958.1993.tb00980.x. [DOI] [PubMed] [Google Scholar]
  26. Latterich M., Watson M. D. Isolation and characterization of osmosensitive vacuolar mutants of Saccharomyces cerevisiae. Mol Microbiol. 1991 Oct;5(10):2417–2426. doi: 10.1111/j.1365-2958.1991.tb02087.x. [DOI] [PubMed] [Google Scholar]
  27. Lindquist S. The heat-shock response. Annu Rev Biochem. 1986;55:1151–1191. doi: 10.1146/annurev.bi.55.070186.005443. [DOI] [PubMed] [Google Scholar]
  28. Mager W. H., Ferreira P. M. Stress response of yeast. Biochem J. 1993 Feb 15;290(Pt 1):1–13. doi: 10.1042/bj2900001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mager W. H., Varela J. C. Osmostress response of the yeast Saccharomyces. Mol Microbiol. 1993 Oct;10(2):253–258. [PubMed] [Google Scholar]
  30. Novick P., Botstein D. Phenotypic analysis of temperature-sensitive yeast actin mutants. Cell. 1985 Feb;40(2):405–416. doi: 10.1016/0092-8674(85)90154-0. [DOI] [PubMed] [Google Scholar]
  31. Otto J., Argos P., Rossmann M. G. Prediction of secondary structural elements in glycerol-3-phosphate dehydrogenase by comparison with other dehydrogenases. Eur J Biochem. 1980 Aug;109(2):325–330. doi: 10.1111/j.1432-1033.1980.tb04798.x. [DOI] [PubMed] [Google Scholar]
  32. Pavlik P., Simon M., Schuster T., Ruis H. The glycerol kinase (GUT1) gene of Saccharomyces cerevisiae: cloning and characterization. Curr Genet. 1993 Jul-Aug;24(1-2):21–25. doi: 10.1007/BF00324660. [DOI] [PubMed] [Google Scholar]
  33. Pidoux A. L., Fawell E. H., Armstrong J. Glycerol-3-phosphate dehydrogenase homologue from Schizosaccharomyces pombe. Nucleic Acids Res. 1990 Dec 11;18(23):7145–7145. doi: 10.1093/nar/18.23.7145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Piper P. W. Molecular events associated with acquisition of heat tolerance by the yeast Saccharomyces cerevisiae. FEMS Microbiol Rev. 1993 Aug;11(4):339–355. doi: 10.1111/j.1574-6976.1993.tb00005.x. [DOI] [PubMed] [Google Scholar]
  35. Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
  36. Rønnow B., Kielland-Brandt M. C. GUT2, a gene for mitochondrial glycerol 3-phosphate dehydrogenase of Saccharomyces cerevisiae. Yeast. 1993 Oct;9(10):1121–1130. doi: 10.1002/yea.320091013. [DOI] [PubMed] [Google Scholar]
  37. Schaaff I., Green J. B., Gozalbo D., Hohmann S. A deletion of the PDC1 gene for pyruvate decarboxylase of yeast causes a different phenotype than previously isolated point mutations. Curr Genet. 1989 Feb;15(2):75–81. doi: 10.1007/BF00435452. [DOI] [PubMed] [Google Scholar]
  38. Sengstag C., Hinnen A. The sequence of the Saccharomyces cerevisiae gene PHO2 codes for a regulatory protein with unusual aminoacid composition. Nucleic Acids Res. 1987 Jan 12;15(1):233–246. doi: 10.1093/nar/15.1.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sharp P. M., Cowe E. Synonymous codon usage in Saccharomyces cerevisiae. Yeast. 1991 Oct;7(7):657–678. doi: 10.1002/yea.320070702. [DOI] [PubMed] [Google Scholar]
  40. Sleep D., Ogden J. E., Roberts N. A., Goodey A. R. Cloning and characterisation of the Saccharomyces cerevisiae glycerol-3-phosphate dehydrogenase (GUT2) promoter. Gene. 1991 May 15;101(1):89–96. doi: 10.1016/0378-1119(91)90228-4. [DOI] [PubMed] [Google Scholar]
  41. Sprague G. F., Cronan J. E. Isolation and characterization of Saccharomyces cerevisiae mutants defective in glycerol catabolism. J Bacteriol. 1977 Mar;129(3):1335–1342. doi: 10.1128/jb.129.3.1335-1342.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Stateva L. I., Oliver S. G., Trueman L. J., Venkov P. V. Cloning and characterization of a gene which determines osmotic stability in Saccharomyces cerevisiae. Mol Cell Biol. 1991 Aug;11(8):4235–4243. doi: 10.1128/mcb.11.8.4235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Thomas B. J., Rothstein R. Elevated recombination rates in transcriptionally active DNA. Cell. 1989 Feb 24;56(4):619–630. doi: 10.1016/0092-8674(89)90584-9. [DOI] [PubMed] [Google Scholar]
  44. Van Aelst L., Hohmann S., Bulaya B., de Koning W., Sierkstra L., Neves M. J., Luyten K., Alijo R., Ramos J., Coccetti P. Molecular cloning of a gene involved in glucose sensing in the yeast Saccharomyces cerevisiae. Mol Microbiol. 1993 May;8(5):927–943. doi: 10.1111/j.1365-2958.1993.tb01638.x. [DOI] [PubMed] [Google Scholar]
  45. Van Aelst L., Hohmann S., Zimmermann F. K., Jans A. W., Thevelein J. M. A yeast homologue of the bovine lens fibre MIP gene family complements the growth defect of a Saccharomyces cerevisiae mutant on fermentable sugars but not its defect in glucose-induced RAS-mediated cAMP signalling. EMBO J. 1991 Aug;10(8):2095–2104. doi: 10.1002/j.1460-2075.1991.tb07742.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Varela J. C., van Beekvelt C., Planta R. J., Mager W. H. Osmostress-induced changes in yeast gene expression. Mol Microbiol. 1992 Aug;6(15):2183–2190. doi: 10.1111/j.1365-2958.1992.tb01392.x. [DOI] [PubMed] [Google Scholar]
  47. Vuorio O. E., Kalkkinen N., Londesborough J. Cloning of two related genes encoding the 56-kDa and 123-kDa subunits of trehalose synthase from the yeast Saccharomyces cerevisiae. Eur J Biochem. 1993 Sep 15;216(3):849–861. doi: 10.1111/j.1432-1033.1993.tb18207.x. [DOI] [PubMed] [Google Scholar]
  48. Wiemken A. Trehalose in yeast, stress protectant rather than reserve carbohydrate. Antonie Van Leeuwenhoek. 1990 Oct;58(3):209–217. doi: 10.1007/BF00548935. [DOI] [PubMed] [Google Scholar]
  49. Yancey P. H., Clark M. E., Hand S. C., Bowlus R. D., Somero G. N. Living with water stress: evolution of osmolyte systems. Science. 1982 Sep 24;217(4566):1214–1222. doi: 10.1126/science.7112124. [DOI] [PubMed] [Google Scholar]
  50. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  51. von Kalm L., Weaver J., DeMarco J., MacIntyre R. J., Sullivan D. T. Structural characterization of the alpha-glycerol-3-phosphate dehydrogenase-encoding gene of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5020–5024. doi: 10.1073/pnas.86.13.5020. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES