Molecular and General Genetics MGG

, Volume 236, Issue 2–3, pp 214–218 | Cite as

Gene SNQ2 of Saccharomyces cerevislae, which confers resistance to 4-nitroquinoline-N-oxide and other chemicals, encodes a 169 kDa protein homologous to ATP-dependent permeases

  • Jörg Servos
  • Eckard Haase
  • Martin Brendel
Article

Summary

The yeast gene SNQ2 confers hyper-resistance to the mutagens 4-nitroquinoline-N-oxide (4-NQO) and Triaziquone, as well as to the chemicals sulphomethuron methyl and phenanthroline when present in multiple copies in transformants of Saccharomyces cerevisiae. Subcloning and sequencing of a 5.5 kb yeast DNA fragment revealed that SNQ2 has an open reading frame of 4.5 kb. The putative encoded polypeptide of 1501 amino acids has a predicted molecular weight of 169 kDa and has several hydrophobic regions. Northern analysis showed a transcript of 5.5 kb. Haploid cells with a disrupted SNQ2 reading frame are viable. The SNQ2-encoded protein has domains believed to be involved in ATP binding and is likely to be membrane associated. It most probably serves as an ATP-dependent permease.

Key words

Mutagen hyper-resistance 4-nitroquinolineN-oxide Yeast ATP-dependent permease 

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References

  1. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1988) Current protocols in molecular biology, vols. 1 & 2, chapters 2 and 13. Wiley Interscience, New YorkGoogle Scholar
  2. Birnboim HG, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523Google Scholar
  3. Chen C, Chin JE, Ueda K, Clark DP, Pastan I, Gottesman MM, Roninson IB (1986) Internal duplication and homology with bacterial transport proteins in the mdr1 (P-Glycoprotein) gene from multidrug-resistant human cells. Cell 47:381–389Google Scholar
  4. Dagert M, Ehrlich SD (1979) Prolonged incubation in calcium chloride improves the competence of E. coli cells. Gene 6:23–28Google Scholar
  5. Del Sal G, Manfioletti G, Schneider C (1988) A one-tube plasmid DNA mini-preparation suitable for sequencing. Nucleic Acids Res 16:9878Google Scholar
  6. Doolittle RF, Johnson MS, Hussain J, Van Houten B, Thomas DC, Sancar A (1986) Domiainal evolution of a prokaryotic DNA repair protein and its relationship to active-transport proteins. Nature 323:451–453Google Scholar
  7. Dreesen TD, Johnson DH, Henikoff S (1988) The brown protein of Drosophila melanogaster is similar to the white protein and to components of active transport complexes. Mol Cell Biol 8:5206–5215Google Scholar
  8. Fleer R, Brendel M (1979) Formation and fate of crosslinks induced by polyfunctional anticancer drugs in yeast. Mol Gen Genet 176:41–52Google Scholar
  9. Gömpel-Klein P, Brendel M (1990) Allelism of SNQ1 and ATR1, genes of the yeast Saccharomyces cerevisiae required for controlling sensitivity to 4-nitroquinoline-N-oxide and aminotriazole. Curr Genet 18:93–96Google Scholar
  10. Gömpel-Klein P, Mack M, Brendel M (1989) Molecular characterization of two genes SNQ and SFA that confer hyperresistance of 4-nitroquinoline-N-oxide and formaldehyde in Saccharomyces cerevisiae. Curr Genet 16:65–74Google Scholar
  11. Grey M, Brendel M (1992) Rapid and simple isolation of DNA from agarose gels. Curr Genet 22:83–84Google Scholar
  12. 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 21:319–324Google Scholar
  13. Henikoff S (1987) Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol 15:156–165Google Scholar
  14. Hertle K, Haase E, Brendel M (1991) The SNQ3 gene of Saccharomyces cerevisiae confers hyper-resistance to several functionally unrelated chemicals. Curr Genet 19:429–433Google Scholar
  15. Higgins CF, Hyde SC, Mimmack MM, Gileadi U, Gill DR, Gallagher MP (1990) Binding protein-dependent transport systems. J Bioenerg Biomembr 22:571–592Google Scholar
  16. Hill JE, Myers AM, Koerner TJ, Tzagoloff A (1986) Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast 2:163–167Google Scholar
  17. Hussain M, Lenard J (1991) Characterization of PDR4, a Saccharomyces cerevisiae gene that confers pleiotropic drug resistance in high-copy number. Gene 101:149–152Google Scholar
  18. Hyde SC, Emsley P, Hartshorn MJ, Mimmack MM, Gileadi U, Pearce SR, Gallagher MP, Gill DR, Hubbard RE, Higgins CF (1990) Structural model of ATP-binding proteins associated with cystic fibrosis, multidrug resistance and bacterial transport. Nature 346:362–365Google Scholar
  19. Ito H, Fukuda J, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163Google Scholar
  20. Kanazawa S, Driscoll M, Struhl K (1988) ATR1, a Saccharomyces cerevisiae gene encoding a transmembrane protein required for aminotriazole resistance. Mol Cell Biol 8:664–673Google Scholar
  21. Kane SE, Pastan I, Gottesman MM (1990) Genetic basis of multidrug resistance of tumor cells. J Bioenerg Biomembr 22:593–618Google Scholar
  22. Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132Google Scholar
  23. Leppert G, McDevitt R, Falco SC, Van Dyke TK, Ficke MB, Golin J (1990) Cloning by gene amplification of two loci conferring multiple drug resistance in Saccharomyces. Genetics 125:13–20Google Scholar
  24. McGrath JP, Varshavsky A (1989) The yeast STE6 gene encodes a homologue of the mammalian multidrug resistance P-glycoprotein. Nature 340:400–404Google Scholar
  25. Moye-Rowley WS, Harshman KD, Parker CS (1989) Yeast YAP1 encodes a novel form of the jun family of transcriptional activator proteins. Genes Dev 3:283–292Google Scholar
  26. Rao MJK, Argos P (1986) A conformational preference parameter to predict helices in integral membrane proteins. Biochim Biophys Acta 869:197–214Google Scholar
  27. Pepling M, Mount MS (1990) Sequence of a cDNA from the Drosophila melanogaster white gene. Nucleic Acids Res 18:1633Google Scholar
  28. Rechsteiner M (1988) Regulation of enzyme levels by proteolysis: the role of PEST regions. Adv Enzyme Regul 27:135–151Google Scholar
  29. Rechsteiner M, Rogers S, Rote K (1987) Protein structure and intracellular stability. Trends Biochem Sci 12:390–394Google Scholar
  30. Riordan JR, Rommens JM, Kremm B-S, Alon N, Rozmahel R, Grzelczak Z, Zielenski J, Lok S, Plavsic N, Chou J-L, Drumm ML, Iannuzzi MC, Collins FS, Tsui L-C (1989) Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 245:1066–1072Google Scholar
  31. Rodriguez RL, Tait RC (1983) Recombinant DNA techniques: An introduction. Addison-Wesley, LondonGoogle Scholar
  32. Rogers S, Wells R, Rechsteiner M (1986) Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science 234:364–368Google Scholar
  33. Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–209Google Scholar
  34. Ruhland AR, Brendel M (1979) Mutagenesis by cytostatic alkylating agents in yeast strains of differing repair capacities. Genetics 92:83–97Google Scholar
  35. Ruhland AR, Haase E, Siede W, Brendel M (1981) Isolation of yeast mutants sensitive to the bifunctional alkylating agent nitrogen mustard. Mol Gen Genet 181:346–351Google Scholar
  36. Sandbaken M, Lupisella JA, DiDomenico B, Chakraburtty K (1990) Isolation and characterization of the structural gene encoding elongation factor 3. Biochim Biophys Acta 1050:230–234Google Scholar
  37. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467Google Scholar
  38. Schnell N, Entian K-D (1991) Identification and characterization of a Saccharomyces cerevisiae gene (PAR1) conferring resistance to iron chelators. Eur J Biochem 200:487–493Google Scholar
  39. Schnell N, Krems B, Entian K-D (1992) The PAR1 (YAP1) gene of Saccharomyces cerevisiae, a c-jun homologue, is involved in oxygen metabolism. Curr Genet 21:269–273Google Scholar
  40. Sharp PM, Li WH (1987) The codon adaptation index — A measure of directional synonymous codon usage bias, and its potential applications. Nucleic Acids Res 15:1281–1295Google Scholar

Copyright information

© Springer-Vertag 1993

Authors and Affiliations

  • Jörg Servos
    • 1
  • Eckard Haase
    • 1
  • Martin Brendel
    • 1
  1. 1.Institut für Mikrobiologie der J.W. Goethe-UniversitätFrankfurt/MainGermany

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