Summary
In the yeast Saccharomyces cerevisiae, the pma1 mutations confers vanadate-resistance to H+-ATPase activity when measured in isolated plasma membranes. In vivo, the growth of pma1 mutants is resistant to Dio-9, ethidium bromide and guanidine derivatives. This phenotype was used to man the pma1 mutation adjacent to LEU1 gene on chromosome VII. From a cosmid library of a wild-type Saccharomyces cerevisiae genome, a large 30 kb DNA fragment was isolated by complementation of a leu1-pma1 double mutant. A 5 kb HindIII fragment was subcloned and it restored both Leu+ and Pma+ phenotypes after integrative transformation. The restriction map of the 5 kb HindIII fragment and Southern blot analysis reveal that the cloned fragment contains the entire structural gene for the plasma membrane ATPase and the 5′ end of the adjacent LEU1 gene. The pma1 mutation conferring vanadate-resistance is thus located in the structural gene for the plasma membrane ATPase.
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References
Bach M, Lacroute F (1972) Direct selective techniques for the isolation of pyrimidine auxotrophs in yeast. Mol Gen Genet 115:126–130
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Clarke L, Carbon J (1980) Isolation of the centromere-linked CDC 10 gene by complementation in yeast. Proc Natl Acad Sci USA 77:2173–2177
Davis RW, Thomas M, Cameron J, St John TP, Scherer S, Padgett RA (1980) Rapid DNA isolation for enzymatic hybridization analysis. Methods Enzymol 65:404–411
Dieckmann CD, Tzagoloff A (1983) Transformation of nuclear respiratory deficient mutants of yeast. Methods Enzymol 97:355–360
Dufour JP, Boutry M, Goffeau A (1980) Plasma membrane AT-Pase of yeast. Comparative inhibition studies of the purified and membrane-bound enzymes. J Biol Chem 255:5735–5741
Falco SC, Yuyang L, Broach JR, Botstein D (1982) Genetic properties of chromosomally integrated 2 μ DNA in yeast. Cell 29:573–584
Falco SC, Rose M, Botstein D (1983) Homologous recombination between episomal plasmids and chromosomes in yeast. Genetics 105:843–856
Foury F, Goffeau A (1975) Stimulation of active uptake of nucleosides and amino acids by cyclic adenosine 3′:5′-monophosphate in the yeast Schizosaccharomyces pombe. J Biol Chem 250:2354–2362
Gerbaud C, Elmerich C, Tandeau de Marsak N, Chocat P, Charpin N, Guérineau M, Aubert JP (1981) Construction of new yeast vectors and cloning of the Nif (nitrogen fixation) gene cluster of Klebsiella pneumoniae in yeast. Curr Genet 3:173–180
Gieldanowski J, Konieczny M, Teodorczyk J, Bierowska D (1971) New bisaminoguanidine derivatives-potential immunodepressive agents. Dissert Pharm Pharmacol 23:335–351
Goffeau A, Slayman CW (1981) The proton-translocating ATPase of the fungal plasma membrane. Biochim Biophys Acta 639:197–223
Goffeau A, Ulaszewski S, Dufour JP, Ghislain M (1984) Properties of plasma membrane ATPase mutants in yeast. Third European Bioenergetics Conference, Short Reports 3B:601–602
Guerineau M, Slonimski P, Avner PR (1974) Yeast episome: oligomycin resistance associated with a small covalently closed nonmitochondrial circular DNA. Biochem Biophys Res Commun 61:462–469
Hieter P, Pridmore D, Hegemann JH, Thomas M, Davis RW, Philipsen P (1985) Functional selection and analysis of yeast centromeric DNA. Cell 42:913–921
Hsiao CL, Carbon J (1981) Direct selection procedure for the isolation of functional centromeric DNA. Proc Natl Acad Sci USA 78:3760–3764
Hsu YP, Schimmel P (1984) Yeast LEU1. Repression of mRNA levels by leucine and relationship of 5′ noncoding region to that of LEU2. J Biol Chem 259:3714–3719
Jackson JA, Fink GR (1981) Gene conversion between duplicated genetic elements in yeast. Nature 292:306–311
Kushner SR (1978) An improved method for transformation of Escherichia coli with ColE1-derived plasmids. In: Boyer HB, Nicosia S (eds) Genetic engineering. Elsevier/North-Holland, Amsterdam, pp 17–28
Lachowicz TM, Konieczny M, Witkowska R (1974) Non-chromosomal respiratory deficient mutants induced by N,N′-(p-xylylidene)-bis-aminoguanidine-2HCl. Acta Microbiol Pol 6:147–154
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Maniatis T, Fritsch EF, Sambrook J (1982) In: Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Mortimer RK, Hawthorne DC (1975) Genetic map** in yeast. Methods Cell Biol 11:221–233
Mortimer RK, Schild D (1980) Genetic map of Saccharomyces cerevisiae. Microbiol Rev 44:519–571
Pullman ME, Penefsky HS (1976) In: Colowick SP, Caplan NO (eds) Methods in enzymology. Academic Press, New York, London, pp 277–284
Rank GH, Bech-Hansen NT (1973) Single nuclear gene inherited cross resistance and collateral sensitivity to 17 inhibitors of mitochondrial function in S. cerevisiae. Mol Gen Genet 126:93–102
Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Roon RJ, Larimore FS, Meyer GM, Kreisle RA (1978) Characterezation of a Dio-9-resistant strain of Saccharomyces cerevisiae. Arch Biochem Biophys 185:142–150
Saunders GW, Rank GH (1982) Allelism of pleiotropic drug resistance in Saccharomyces cerevisiee. Can J Genet Cytol 24:493–503
Serrano R, Kielland-Brandt MC, Fink GR (1986) Yeast plasma membrane ATPase is essential for growth and has homology with (Na+ +K+), K+- and Ca2+-ATPases. Nature 319:686–693
Snow R (1979) Maximum likelihood estimation of linkage and inheritance from tetrad data. Genetics 92:231–245
Šubik J, Ulaszewski S, Goffeau A (1986) Genetic map** of nuclear mucidin resistance mutations in Saccharomyces cerevisiae. A new pdr locus on chromosome II. Curr Genet 10:665–670
Ulaszewski S, Grenson M, Goffeau A (1983) Modified plasmamembrane ATPase in mutants of Saccharomyces cerevisiae. Eur J Biochem 130:235–239
Ulaszewski S, Coddington A, Goffeau A (1986a) A new mutation for multiple drug resistance and modified plasma membrane ATPase activity in Schizosaccharomyces pombe. Curr Genet 10:359–364
Ulaszewski S, Van Herck JC, Dufour JP, Kulpa J, Nieuwenhuis B, Goffeau A (1987) A single mutation confers vanadate resistance to the plasma membrane H+-ATPase from the yeast Schizosaccharomyces pombe. J Biol Chem 262:223–228
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Communicated by C.P. Hollenberg
Publication no 2456 from the Biology Directorate of the Commission of European Communities
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Ulaszewski, S., Balzi, E. & Goffeau, A. Genetic and molecular map** of the pma1 mutation conferring vanadate resistance to the plasma membrane ATPase from Saccharomyces cerevisiae . Mol Gen Genet 207, 38–46 (1987). https://doi.org/10.1007/BF00331488
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DOI: https://doi.org/10.1007/BF00331488