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The Saccharomyces cerevisiae SGE1 gene product: a novel drug-resistance protein within the major facilitator superfamily

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Abstract

Several pleiotropic drug sensitivities have been described in yeast. Some involve the loss of putative drug efflux pumps analogous to mammalian P-glycoproteins, others are caused by defects in sterol synthesis resulting in higher plasma membrane permeability. We have constructed a Saccharomyces cerevisiae strain that exhibits a strong crystal violet-sensitive phenotype. By selecting cells of the supersensitive strain for normal sensitivity after transformation with a wild-type yeast genomic library, a complementing 10-kb DNA fragment was isolated, a 3.4-kb subfragment of which was sufficient for complementation. DNA sequence analysis revealed that the complementing fragment comprised the recently sequenced SGE1 gene, a partial multicopy suppressor of gal11 mutations. The supersensitive strain was found to be a sge1 null mutant. Overexpression of SGE1 on a high-copy-number plasmid increased the resistance of the supersensitive strain. Disruption of SGE1 in a wild-type strain increased the sensitivity of the strain. These features of the SGE1 phenotype, as well as sequence homologies of SGE1 at the amino acid level, confirm that the Sge1 protein is a member of the drug-resistance protein family within the major facilitator superfamily (MFS).

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References

  • Amakasu H, Suzuki Y, Nishizawa M, Fukasawa T (1993) Isolation and characterization of SGE1: a yeast gene that partially suppresses the gal11 mutation in multiple copies. Genetics 134:675–683

    Google Scholar 

  • Ausubel F, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1990) Current protocols in molecular biology. John Wiley & Sons, New York, NY

    Google Scholar 

  • Bard M, Lees ND, Burrows LS, Kleinhans FW (1978) Differences in crystal violet uptake and cation-induced death among yeast sterol mutants. J Bacteriol 135:1146–1148

    Google Scholar 

  • Bennetzen JL, Hall BD (1982) Codon selection in yeast. J Biol Chem 257:3026–3031

    Google Scholar 

  • Bilofsky HS, Burks C (1988) The GenBank genetic sequence data bank. Nucleic Acids Res 16:1861–1863

    Google Scholar 

  • Chen WN, Capieaux E, Balzi E, Goffeau A (1991) The YGL022 gene encodes a putative transport protein. Yeast 7:305–308

    Google Scholar 

  • Coque JJ, Liras P, Martin JF (1993) Genes for a β-lactamase, a penicillin-binding protein and a transmembrane protein are clustered with the cephamycin biosynthetic genes in Nocardia lactamdurans. EMBO J 12:631–639

    Google Scholar 

  • De-Nobel JG, Barnett JA (1991) Passage of molecules through yeast cell walls: a brief essay-review. Yeast 7:313–323

    Google Scholar 

  • De-Nobel JG, Klis FM, Munnik T, Priem J, van-den-Ende H (1990) An assay of relative cell wall porosity in Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe. Yeast 6:483–490

    Google Scholar 

  • Fassler JS, Winston F (1989) The Saccharomyces cerevisiae SPT13/GAL11 gene has both positive and negative regulatory roles in transcription. Mol Cell Biol 9:5602–5609

    Google Scholar 

  • Felix H (1982) Permeabilized cells. Anal Biochem 120:211–234

    Google Scholar 

  • Goffeau A, Nakai K, Slorninski P, Risler JL (1993) The membrane proteins encoded by yeast chromosome III genes. FEBS Lett 325:112–117

    Google Scholar 

  • Gottesman MM, Pastan I (1988) The multidrug transporter, a double-edged sword. J Biol Chem 263:12163–12166

    Google Scholar 

  • Graham TR, Scott PA, Emr SD (1993) Brefeldin A reversibly blocks early but not late protein transport steps in the yeast secretory pathway. EMBO J 12:869–877

    Google Scholar 

  • Guthrie C, Fink GR (1991) Guide to yeast genetics and molecular biology. Academic Press, San Diego

    Google Scholar 

  • Haase E, Servos J, Brendel M (1992) Isolation and characterization of additional genes influencing resistance to various mutagens in the yeast Saccharomyces cerevisiae. Curr Genet 21:319–324

    Google Scholar 

  • Hill JE, Myers AM, Koerner TJ, Tzagoloff A (1986) Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast 2:163–167

    Google Scholar 

  • Hoffman CS, Winston F (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57:267–272

    Google Scholar 

  • Klebe RJ, Harriss JV, Sharp ZD, Douglas MG (1983) A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast. Gene 25:333–341

    Google Scholar 

  • Kuchler K, Sterne RE, Thorner J (1989) Saccharomyces cerevisiae STE6 gene product: a novel pathway for protein export in eukaryotic cells. EMBO J 8:3973–3984

    Google Scholar 

  • Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132

    CAS  PubMed  Google Scholar 

  • Lehninger AL (1977) Biochemistry. Worth Publishers, New York

    Google Scholar 

  • Leppert G, McDevitt R, Falco SC, Van-Dyk TK, Ficke MB, Golin J (1990) Cloning by gene amplification of two loci conferring multiple drug resistance in Saccharomyces. Genetics 125:13–20

    Google Scholar 

  • Marger MD, Saier M Jr (1993) A major superfamily of transmembrane facilitators that catalyse uniport, symport and antiport. Trends Biochem Sci 18:13–20

    Google Scholar 

  • McCammon MT, Hartmann MA, Bottema CD, Parks LW (1984) Sterol methylation in Saccharomyces cerevisiae. J Bacteriol 157:475–483

    Google Scholar 

  • McGrath JP, Varshavsky A (1989) The yeast STE6 gene encodes a homologue of the mammalian multidrug resistance P-glycoprotein. Nature 340:400–404

    Google Scholar 

  • Meyers S, Schauer W, Balzi E, Wagner M, Goffeau A, Golin J (1992) Interaction of the yeast pleiotropic drug resistance genes PDR1 and PDR5. Curr Genet 21:431–436

    Google Scholar 

  • Morita T, Yanagihara Y (1985) Osmotic-sensitive mutants of Saccharomyces cerevisiae as screening organisms for promutagens and procarcinogens. Chem Pharm Bull Tokyo 33:1576–1582

    Google Scholar 

  • Nishizawa M, Suzuki Y, Nogi Y, Matsumoto K, Fukasawa T (1990) Yeast Gal11 protein mediates the transcriptional activation signal of two different transacting factors, Gal4 and general regulatory factor I/repressor/activator site binding protein 1/translation upstream factor. Proc Natl Acad Sci USA 87:5373–5377

    Google Scholar 

  • Nitiss J, Wang JC (1988) DNA topoisomerase-targeting antitumor drugs can be studied in yeast. Proc Natl Acad Sci USA 85:7501–7505

    Google Scholar 

  • Peden KW (1983) Revised sequence of the tetracycline-resistance gene of pBR322. Gene 22:277–280

    Google Scholar 

  • Rank GH, Robertson A, Phillips K (1975) Reduced plasma membrane permeability in a multiple cross-resistant strain of Saccharomyces cerevisiae. J Bacteriol 122:359–366

    Google Scholar 

  • Riordan JR, Rommens JM, Kerem B, Alon N, Rozmahel R, Grzelczak Z, Zielenski J, Lok S, Plavsic N, Chou JL (1989) Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 245:1066–1073

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sauer B (1992) Identification of cryptic lox sites in the yeast genome by selection for Cre-mediated chromosome translocations that confer multiple drug resistance. J Mol Biol 223:911–928

    Google Scholar 

  • Scherrer R, Louden L, Gerhardt P (1974) Porosity of the yeast cell wall and membrane. J Bacteriol 118:534–540

    Google Scholar 

  • Sengstag C (1993) The sequence of Saccharomyces cerevisiae cloning vector pCS19 following direct selection for DNA inserts. Gene 124:141–142

    Google Scholar 

  • Sengstag C, Hinnen A (1987) The sequence of the Saccharomyces cerevisiae gene PH02 codes for a regulatory protein with unusual amino acid composition. Nucleic Acids Res 15:233–246

    Google Scholar 

  • Servos J, Haase E, Brendel M (1993) Gene SNQ2 of Saccharomyces cerevisiae, which confers resistance to 4-nitroquinoline-N-oxide and other chemicals, encodes a 169 kDa protein homologous to ATP-dependent permeases. Mol Gen Genet 236:214–218

    Google Scholar 

  • Sherman F (1991) Getting started with yeast. Methods Enzymol 194:3–21

    Google Scholar 

  • Sikorski RS, Hieter P (1989) A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122:19–27

    CAS  PubMed  Google Scholar 

  • Suzuki Y, Nogi Y, Abe A, Fukasawa T (1988) GAL11 protein, an auxiliary transcription activator for genes encoding galactose-metabolizing enzymes in Saccharomyces cerevisiae. Mol Cell Biol 8:4991–4999

    Google Scholar 

  • Ulaszewski S, Balzi E, Goffeau A (1987) Genetic and molecular mapping of the pma1 mutation conferring vanadate resistance to the plasma membrane ATPase from Saccharomyces cerevisiae. Mol Gen Genet 207:38–46

    Google Scholar 

  • Wang M, Balzi E, Van Dyck L, Golin J, Goffeau A (1992) Sequencing of the yeast multidrug resistance PDR5 gene encoding a putative pump for drug efflux. Yeast 8:S528

    Google Scholar 

  • Winsor B, Potter AA, Karst F, Nestmann ER, Lacroute F (1987) Characterization of a yeast mutation ise1 that enhances permeability of Saccharomyces cerevisiae. Environ Mutagen 9:114

    Google Scholar 

  • Zimmermann FK, Kern R, Rasenberger H (1975) A yeast strain for simultaneous detection of induced mitotic crossing over, mitotic gene conversion and reverse mutation. Mutat Res 28:381–388

    Google Scholar 

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Authors and Affiliations

  1. Institute of Toxicology, Swiss Federal Institute of Technology and University of Zürich, Schorenstrasse 16, 8603, Schwerzenbach, Switzerland

    Ann E. Ehrenhofer-Murray, Friedrich E. Würgler & Christian Sengstag

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  1. Ann E. Ehrenhofer-Murray
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  2. Friedrich E. Würgler
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  3. Christian Sengstag
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Communicated by C. P. Hollenberg

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Ehrenhofer-Murray, A.E., Würgler, F.E. & Sengstag, C. The Saccharomyces cerevisiae SGE1 gene product: a novel drug-resistance protein within the major facilitator superfamily. Molec. Gen. Genet. 244, 287–294 (1994). https://doi.org/10.1007/BF00285456

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  • DOI: https://doi.org/10.1007/BF00285456

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