Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1998 Aug 3;17(15):4257–4265. doi: 10.1093/emboj/17.15.4257

The pdr12 ABC transporter is required for the development of weak organic acid resistance in yeast.

P Piper 1, Y Mahé 1, S Thompson 1, R Pandjaitan 1, C Holyoak 1, R Egner 1, M Mühlbauer 1, P Coote 1, K Kuchler 1
PMCID: PMC1170759  PMID: 9687494

Abstract

Exposure of Saccharomyces cerevisiae to sorbic acid strongly induces two plasma membrane proteins, one of which is identified in this study as the ATP-binding cassette (ABC) transporter Pdr12. In the absence of weak acid stress, yeast cells grown at pH 7.0 express extremely low Pdr12 levels. However, sorbate treatment causes a dramatic induction of Pdr12 in the plasma membrane. Pdr12 is essential for the adaptation of yeast to growth under weak acid stress, since Deltapdr12 mutants are hypersensitive at low pH to the food preservatives sorbic, benzoic and propionic acids, as well as high acetate levels. Moreover, active benzoate efflux is severely impaired in Deltapdr12 cells. Hence, Pdr12 confers weak acid resistance by mediating energy-dependent extrusion of water-soluble carboxylate anions. The normal physiological function of Pdr12 is perhaps to protect against the potential toxicity of weak organic acids secreted by competitor organisms, acids that will accumulate to inhibitory levels in cells at low pH. This is the first demonstration that regulated expression of a eukaryotic ABC transporter mediates weak organic acid resistance development, the cause of widespread food spoilage by yeasts. The data also have important biotechnological implications, as they suggest that the inhibition of this transporter could be a strategy for preventing food spoilage.

Full Text

The Full Text of this article is available as a PDF (364.9 KB).

Selected References

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

  1. Balzi E., Chen W., Ulaszewski S., Capieaux E., Goffeau A. The multidrug resistance gene PDR1 from Saccharomyces cerevisiae. J Biol Chem. 1987 Dec 15;262(35):16871–16879. [PubMed] [Google Scholar]
  2. Balzi E., Wang M., Leterme S., Van Dyck L., Goffeau A. PDR5, a novel yeast multidrug resistance conferring transporter controlled by the transcription regulator PDR1. J Biol Chem. 1994 Jan 21;269(3):2206–2214. [PubMed] [Google Scholar]
  3. Bissinger P. H., Kuchler K. Molecular cloning and expression of the Saccharomyces cerevisiae STS1 gene product. A yeast ABC transporter conferring mycotoxin resistance. J Biol Chem. 1994 Feb 11;269(6):4180–4186. [PubMed] [Google Scholar]
  4. Boeke J. D., Trueheart J., Natsoulis G., Fink G. R. 5-Fluoroorotic acid as a selective agent in yeast molecular genetics. Methods Enzymol. 1987;154:164–175. doi: 10.1016/0076-6879(87)54076-9. [DOI] [PubMed] [Google Scholar]
  5. Bolard J. How do the polyene macrolide antibiotics affect the cellular membrane properties? Biochim Biophys Acta. 1986 Dec 22;864(3-4):257–304. doi: 10.1016/0304-4157(86)90002-x. [DOI] [PubMed] [Google Scholar]
  6. Casal M., Cardoso H., Leão C. Mechanisms regulating the transport of acetic acid in Saccharomyces cerevisiae. Microbiology. 1996 Jun;142(Pt 6):1385–1390. doi: 10.1099/13500872-142-6-1385. [DOI] [PubMed] [Google Scholar]
  7. Cheng L., Watt R., Piper P. W. Polyubiquitin gene expression contributes to oxidative stress resistance in respiratory yeast (Saccharomyces cerevisiae). Mol Gen Genet. 1994 May 10;243(3):358–362. doi: 10.1007/BF00301072. [DOI] [PubMed] [Google Scholar]
  8. Cubitt A. B., Heim R., Adams S. R., Boyd A. E., Gross L. A., Tsien R. Y. Understanding, improving and using green fluorescent proteins. Trends Biochem Sci. 1995 Nov;20(11):448–455. doi: 10.1016/s0968-0004(00)89099-4. [DOI] [PubMed] [Google Scholar]
  9. Decottignies A., Goffeau A. Complete inventory of the yeast ABC proteins. Nat Genet. 1997 Feb;15(2):137–145. doi: 10.1038/ng0297-137. [DOI] [PubMed] [Google Scholar]
  10. Decottignies A., Lambert L., Catty P., Degand H., Epping E. A., Moye-Rowley W. S., Balzi E., Goffeau A. Identification and characterization of SNQ2, a new multidrug ATP binding cassette transporter of the yeast plasma membrane. J Biol Chem. 1995 Jul 28;270(30):18150–18157. doi: 10.1074/jbc.270.30.18150. [DOI] [PubMed] [Google Scholar]
  11. Delahodde A., Delaveau T., Jacq C. Positive autoregulation of the yeast transcription factor Pdr3p, which is involved in control of drug resistance. Mol Cell Biol. 1995 Aug;15(8):4043–4051. doi: 10.1128/mcb.15.8.4043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Delaveau T., Delahodde A., Carvajal E., Subik J., Jacq C. PDR3, a new yeast regulatory gene, is homologous to PDR1 and controls the multidrug resistance phenomenon. Mol Gen Genet. 1994 Sep 1;244(5):501–511. doi: 10.1007/BF00583901. [DOI] [PubMed] [Google Scholar]
  13. Deák T. Foodborne yeasts. Adv Appl Microbiol. 1991;36:179–278. doi: 10.1016/s0065-2164(08)70454-4. [DOI] [PubMed] [Google Scholar]
  14. Egner R., Mahé Y., Pandjaitan R., Kuchler K. Endocytosis and vacuolar degradation of the plasma membrane-localized Pdr5 ATP-binding cassette multidrug transporter in Saccharomyces cerevisiae. Mol Cell Biol. 1995 Nov;15(11):5879–5887. doi: 10.1128/mcb.15.11.5879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Egner R., Rosenthal F. E., Kralli A., Sanglard D., Kuchler K. Genetic separation of FK506 susceptibility and drug transport in the yeast Pdr5 ATP-binding cassette multidrug resistance transporter. Mol Biol Cell. 1998 Feb;9(2):523–543. doi: 10.1091/mbc.9.2.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fernandes L., Rodrigues-Pousada C., Struhl K. Yap, a novel family of eight bZIP proteins in Saccharomyces cerevisiae with distinct biological functions. Mol Cell Biol. 1997 Dec;17(12):6982–6993. doi: 10.1128/mcb.17.12.6982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fleet G. Spoilage yeasts. Crit Rev Biotechnol. 1992;12(1-2):1–44. doi: 10.3109/07388559209069186. [DOI] [PubMed] [Google Scholar]
  18. Henriques M., Quintas C., Loureiro-Dias M. C. Extrusion of benzoic acid in Saccharomyces cerevisiae by an energy-dependent mechanism. Microbiology. 1997 Jun;143(Pt 6):1877–1883. doi: 10.1099/00221287-143-6-1877. [DOI] [PubMed] [Google Scholar]
  19. Katzmann D. J., Hallstrom T. C., Mahé Y., Moye-Rowley W. S. Multiple Pdr1p/Pdr3p binding sites are essential for normal expression of the ATP binding cassette transporter protein-encoding gene PDR5. J Biol Chem. 1996 Sep 20;271(38):23049–23054. doi: 10.1074/jbc.271.38.23049. [DOI] [PubMed] [Google Scholar]
  20. Katzmann D. J., Hallstrom T. C., Voet M., Wysock W., Golin J., Volckaert G., Moye-Rowley W. S. Expression of an ATP-binding cassette transporter-encoding gene (YOR1) is required for oligomycin resistance in Saccharomyces cerevisiae. Mol Cell Biol. 1995 Dec;15(12):6875–6883. doi: 10.1128/mcb.15.12.6875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Krebs H. A., Wiggins D., Stubbs M., Sols A., Bedoya F. Studies on the mechanism of the antifungal action of benzoate. Biochem J. 1983 Sep 15;214(3):657–663. doi: 10.1042/bj2140657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kuchler K., Dohlman H. G., Thorner J. The a-factor transporter (STE6 gene product) and cell polarity in the yeast Saccharomyces cerevisiae. J Cell Biol. 1993 Mar;120(5):1203–1215. doi: 10.1083/jcb.120.5.1203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kuge S., Jones N. YAP1 dependent activation of TRX2 is essential for the response of Saccharomyces cerevisiae to oxidative stress by hydroperoxides. EMBO J. 1994 Feb 1;13(3):655–664. doi: 10.1002/j.1460-2075.1994.tb06304.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Li Z. S., Szczypka M., Lu Y. P., Thiele D. J., Rea P. A. The yeast cadmium factor protein (YCF1) is a vacuolar glutathione S-conjugate pump. J Biol Chem. 1996 Mar 15;271(11):6509–6517. doi: 10.1074/jbc.271.11.6509. [DOI] [PubMed] [Google Scholar]
  25. Mahé Y., Lemoine Y., Kuchler K. The ATP binding cassette transporters Pdr5 and Snq2 of Saccharomyces cerevisiae can mediate transport of steroids in vivo. J Biol Chem. 1996 Oct 11;271(41):25167–25172. doi: 10.1074/jbc.271.41.25167. [DOI] [PubMed] [Google Scholar]
  26. Mahé Y., Parle-McDermott A., Nourani A., Delahodde A., Lamprecht A., Kuchler K. The ATP-binding cassette multidrug transporter Snq2 of Saccharomyces cerevisiae: a novel target for the transcription factors Pdr1 and Pdr3. Mol Microbiol. 1996 Apr;20(1):109–117. doi: 10.1111/j.1365-2958.1996.tb02493.x. [DOI] [PubMed] [Google Scholar]
  27. Martínez-Pastor M. T., Marchler G., Schüller C., Marchler-Bauer A., Ruis H., Estruch F. The Saccharomyces cerevisiae zinc finger proteins Msn2p and Msn4p are required for transcriptional induction through the stress response element (STRE). EMBO J. 1996 May 1;15(9):2227–2235. [PMC free article] [PubMed] [Google Scholar]
  28. Miyahara K., Hirata D., Miyakawa T. yAP-1- and yAP-2-mediated, heat shock-induced transcriptional activation of the multidrug resistance ABC transporter genes in Saccharomyces cerevisiae. Curr Genet. 1996 Jan;29(2):103–105. doi: 10.1007/BF02221572. [DOI] [PubMed] [Google Scholar]
  29. Moye-Rowley W. S., Harshman K. D., Parker C. S. Yeast YAP1 encodes a novel form of the jun family of transcriptional activator proteins. Genes Dev. 1989 Mar;3(3):283–292. doi: 10.1101/gad.3.3.283. [DOI] [PubMed] [Google Scholar]
  30. Piper P. W., Ortiz-Calderon C., Holyoak C., Coote P., Cole M. Hsp30, the integral plasma membrane heat shock protein of Saccharomyces cerevisiae, is a stress-inducible regulator of plasma membrane H(+)-ATPase. Cell Stress Chaperones. 1997 Mar;2(1):12–24. doi: 10.1379/1466-1268(1997)002<0012:htipmh>2.3.co;2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Ruis H., Schüller C. Stress signaling in yeast. Bioessays. 1995 Nov;17(11):959–965. doi: 10.1002/bies.950171109. [DOI] [PubMed] [Google Scholar]
  33. Servos J., Haase E., Brendel M. 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. 1993 Jan;236(2-3):214–218. doi: 10.1007/BF00277115. [DOI] [PubMed] [Google Scholar]
  34. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Stratford M., Anslow P. A. Comparison of the inhibitory action on Saccharomyces cerevisiae of weak-acid preservatives, uncouplers, and medium-chain fatty acids. FEMS Microbiol Lett. 1996 Aug 15;142(1):53–58. doi: 10.1111/j.1574-6968.1996.tb08407.x. [DOI] [PubMed] [Google Scholar]
  36. Szczypka M. S., Wemmie J. A., Moye-Rowley W. S., Thiele D. J. A yeast metal resistance protein similar to human cystic fibrosis transmembrane conductance regulator (CFTR) and multidrug resistance-associated protein. J Biol Chem. 1994 Sep 9;269(36):22853–22857. [PubMed] [Google Scholar]
  37. Wach A., Brachat A., Alberti-Segui C., Rebischung C., Philippsen P. Heterologous HIS3 marker and GFP reporter modules for PCR-targeting in Saccharomyces cerevisiae. Yeast. 1997 Sep 15;13(11):1065–1075. doi: 10.1002/(SICI)1097-0061(19970915)13:11<1065::AID-YEA159>3.0.CO;2-K. [DOI] [PubMed] [Google Scholar]
  38. Wemmie J. A., Szczypka M. S., Thiele D. J., Moye-Rowley W. S. Cadmium tolerance mediated by the yeast AP-1 protein requires the presence of an ATP-binding cassette transporter-encoding gene, YCF1. J Biol Chem. 1994 Dec 23;269(51):32592–32597. [PubMed] [Google Scholar]
  39. Wendler F., Bergler H., Prutej K., Jungwirth H., Zisser G., Kuchler K., Högenauer G. Diazaborine resistance in the yeast Saccharomyces cerevisiae reveals a link between YAP1 and the pleiotropic drug resistance genes PDR1 and PDR3. J Biol Chem. 1997 Oct 24;272(43):27091–27098. doi: 10.1074/jbc.272.43.27091. [DOI] [PubMed] [Google Scholar]
  40. Wolfger H., Mahé Y., Parle-McDermott A., Delahodde A., Kuchler K. The yeast ATP binding cassette (ABC) protein genes PDR10 and PDR15 are novel targets for the Pdr1 and Pdr3 transcriptional regulators. FEBS Lett. 1997 Dec 1;418(3):269–274. doi: 10.1016/s0014-5793(97)01382-3. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES