Abstract
In order to isolate new mutations impairing transcriptional regulation of sulfur metabolism inSaccharomyces cerevisiae, we used a potent genetic screen based on a gene fusion expressing XylE (fromPseudomonas putida) under the control of the promoter region ofMET25. This selection yielded strains mutated in various different genes. We describe in this paper the properties of one of them,MET27. Mutation or disruption ofMET27 leads to a methionine requirement and affects S-adenosylmethionine (AdoMet)-mediated transcriptional control of genes involved in sulfur metabolism. The cloning and sequencing ofMET27 showed that it is identical toVPS33. Disruptions or mutations of geneVPS33 are well known to impair the biogenesis and inheritance of the vacuolar compartment. However, the methionine requirement ofvps33 mutants has not been reported previously. We show here, moreover, that other vps mutants of class C (no apparent vacuoles) also require methionine for growth. Northern blotting experiments revealed that themet27-1 mutation delayed derepression of the transcription of genes involved in sulfur metabolism. By contrast, this delay was not observed in amet27 disrupted strain. Physiological and morphological analyses ofmet27-1 andmet27 disrupted strains showed that these results could be explained by alterations in the ability of the vacuole to transport or store AdoMet, the physiological effector of the transcriptional regulation of sulfur metabolism.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Baker RE, Masison DC (1990) Isolation of the gene encoding theSaccharomyces cerevisiae centromere binding protein CP1. Mol Cell Biol 10:2458–2467
Baldari C, Cesareni G (1985) Plasmids pEMBLY: new single stranded shuttle vectors for the recovery and analysis of yeast DNA sequences. Gene 35:27–32
Banta LM, Vida TA, Herman PK, Emr SD (1990) Characterization of yeast Vps33p, a protein required for vacuolar protein sorting and vacuole biogenesis. Mole Cell Biol 10:4638–4649
Bram RJ, Kornberg RD (1987) Isolation of a centromere DNA binding protein, its homolog, and its possible role as a transcription factor. Mol Cell Biol 7:403–409
Cai M, Davis RW (1990) Yeast centromerc binding protein CBF1, of the helix-loop-helix protein family, is required for chromosome stability and methionine prototrophy. Cell 61:437–446
Cantoni GL (1977) S-Adensylmethionine: present status and future perspectives. In: Salvatore F, Borek E, Zappia V, Williams-Ashmann HG, Schlenk F (eds) The biochemistry of adenosylmethionine. Columbia University Press, New York, pp 557–577
Cherest H, Surdin-Kerjan Y (1992) Genetic analysis of a new mutation conferring cysteine auxotrophy inSaccharomyces cerevisiae: updating of the sulfur metabolism pathway. Genetics 130:51–58
Cherest H, Nguyen Ngoc T, Surdin-Kerjan Y (1985) Transcriptional regulation of theMET3 gene fromSaccharomyces cerevisiae. Gene 34:269–281
Dang CV, Dolde C, Gillison ML, Kato GJ (1992) Discrimination between related DNA sites by a single amino acid residue of Myc related basic-helix-loop-helix proteins. Proc Natl Acad Sci USA 89:599–602
Dessen P, Fondrat C, Valencien C, Mugnier C (1990) Bisance: a French service for access to biomolecular databases. Comput Appl Biosci 6:355–356
Farooqui JZ, Lee HW, Kim S, Paik WK (1983) Studies on compartmentation of S-adenosyl-l-methionine inSaccharomyces cerevisiae and isolated rat hepatocytes. Biophys Biochim Acta 757:342–351
Hieter P, Pridmore D, Hegemann JH, Thomas H, Davis RW, Philippsen P (1985) Functional selective and analysis of yeast centromeric DNA. Cell 42:913–921
Hoffmann CS, Winston F (1987) A ten minutes DNA preparation from yeast releases autonomous plasmids for transformation ofEscherichia coli. Gene 57:267–272
Ito H, Fukuda I, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
Jones EW (1977) Proteinase mutants ofSaccharomyces cerevisiae. Genetics 85:23–33
Kitamoto K, Yoshizawa K, Oshumi Y, Anraku Y (1988) Mutants ofSaccharomyces cerevisiae with defective vacuolar function. Mol Cell Biol 170:2687–2691
Klionsky DJ, Herman PK, Emr SD (1990) The fungal vacuole: composition, function and biogenesis. Microbiol Rev 54:266–292
Köhrer K, Emr SD (1993) The yeastVPS17 gene encodes a membrane-associated protein required for the sorting of soluble vacuolar hydrolases. J Biol Chem 268:559–569
Mellor J, Jiang W, Funk M, Rathjen J, Barnes CA, Hinz T, Hegemann JH, Philippsen P (1990) CPF1, a yeast protein which functions in centromeres and promoters. EMBO J 9:4017–4026
Mellor J, Rathjen J, Jiang W, Dowell SJ (1991) DNA binding of CPF1 is required for optimal centromere function but not for maintaining methionine prototrophy in yeast. Nucleic Acids Res 19:2961–2969
Ohya Y, Oshumi Y, Anraku Y (1986) Isolation and characterization of Ca++ mutants ofSaccharomyces cerevisiae. J Gen Microbiol 132:979–988
Raymond CK, Roberts CJ, Moore KE, Howald I, Stevens TH (1992) Biogenesis of the vacuole inSaccharomyces cerevisiae. Int Rev Cytol 139:59–120
Riva A (1974) A simple and rapid staining method for enhancing the contrast of tissues previously treated with uranyl acetate. J Microsc 19:105–108
Salat-Trepat JM, Evans WC (1971) The meta clearvage of catechol byAzotobacter species. Eur J Biochem 20:400–413
Schwenke J, Robichon-Szulmajster H (1976) The transport of S-adenosyl-l-methionine in isolated yeast vacuoles and spheroplasts. Eur J Biochem 65:49–60
Sikorski RS, Hieter P (1989) A system of shuttle vectors and yeast strains designed for efficient manipulation of DNA inSaccharomyces cerevisiae. Genetics 122:19–27
Shapiro SK, Ehninger DJ (1966) Methods for the analysis and preparation of adenosylhomocysteine. Anal Biochem 15:323–333
Thomas PS (1980) Hybridization of denatured DNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205
Thomas D, Surdin-Kerjan Y (1989) Structure of theHOM2 gene ofSaccharomyces cerevisiae and regulation of its expression. Mol Gen Genet 217:149–154
Thomas D, Surdin-Kerjan Y (1990) An improved strategy for generating a family of unidirectional deletions on large DNA fragments. Gene Anal Tech Appl 7:87–90
Thomas D, Rothstein R, Rosenberg N, Surdin-Kerjan Y (1988) SAM2 encodes the second methionine S-adenosyl transferase inSaccharomyces cerevisiae: physiology and regulation of both enzymes. Mol Cell Biol 8:5132–5139
Thomas D, Cherest H, Surdin-Kerjan Y (1989) Elements involved in S-adenosylmethionine mediated regulation of theSaccharomyces cerevisiaeMET25 gene. Mol Cell Biol 9:3292–3298
Thomas D, Barbey R, Henry D, Surdin-Kerjan Y (1992a) Physiological analysis of mutants ofSaccharomyces cerevisiae impaired in sulphate assimilation. J Gen Microbiol 138:2021–2028
Thomas D, Jacquemin I, Surdin-Kerjan Y (1992b) MET4, a leucine zipper protein, and centromere binding factor I, are both required for transcriptional activation of sulfur metabolism inSaccharomyces cerevisiae. Mol Cell Biol 12:1719–1727
Wada Y, Kitamoto K, Kanbe T, Tanaka K, Anraku Y (1990) TheSLPI gene ofSaccharomyces cerevisiae is essential for vacuolar morphogenesis and function. Mol Cell Biol 10:2214–2223
Wada Y, Oshumi Y, Anraku Y (1992) Genes for directing vacuolar morphogenesis inSaccharomyces cerevisiae. J Biol Chem 267:8665–8670
Worsay MJ, Williams PA (1975) Metabolism of toluene and xylene byPseudomonas putida (arvilla) mt-2: evidence for a new function of the TOL plasmid. J Bacteriol 124:7–13
Wright R, Rine J (1989) Transmission electron microscopy and immunocytochemical studies of yeast: analysis of HMG-CoA reductase overproduction by electron microscopy. Methods Cell Biol 31:472–512
Author information
Authors and Affiliations
Additional information
Communicated by C. Hollenberg I.
Rights and permissions
About this article
Cite this article
Jacquemin-Faure, I., Thomas, D., Laporte, J. et al. The vacuolar compartment is required for sulfur amino acid homeostasis inSaccharomyces cerevisiae . Molec. Gen. Genet. 244, 519–529 (1994). https://doi.org/10.1007/BF00583903
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00583903