European Journal of Plant Pathology

, Volume 108, Issue 7, pp 719–734 | Cite as

Secretion of Natural and Synthetic Toxic Compounds from Filamentous Fungi by Membrane Transporters of the ATP-binding Cassette and Major Facilitator Superfamily

  • Ioannis Stergiopoulos
  • Lute-Harm Zwiers
  • Maarten A. De Waard


This review provides an overview of members of the ATP-binding cassette (ABC) and major facilitator superfamily (MFS) of transporters identified in filamentous fungi. The most common function of these membrane proteins is to provide protection against natural toxic compounds present in the environment of fungi, such as antibiotics produced by other microorganisms. In plant pathogenic fungi, these transporters can also be an important determinant of virulence on host plants by providing protection against plant defence compounds or mediating the secretion of host-specific toxins. Furthermore, they play a critical role in determining base-line sensitivity to fungicides and other antimycotic agents. Overexpression of some of these transporters can lead to the development of resistance to chemically-unrelated compounds, a phenomenon described as multidrug resistance (MDR). This has been observed in a variety of organisms and can impose a serious threat to the effective control of pathogenic fungi.

antibiotic compounds fungicides mycotoxin pathogenicity plant defence compounds resistance host-specific toxin 


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  1. al Shawi MK and Senior AE (1993) Characterization of the adenosine triphosphatase activity of Chinese hamster P-glycoprotein. Journal of Biological Chemistry 268: 4197–4206Google Scholar
  2. Albertson G, Niimi M, Cannon R and Jenkinson H (1996) Multiple efflux mechanisms are involved in Candida albicans fluconazole resistance. Antimicrobial Agents and Chemotherapy 40: 2835–2841Google Scholar
  3. Alexander NJ, McCormick SP and Hohn TM (1999) TRI12, a trichothecene efflux pump from Fusarium sporotrichioides: gene isolation and expression in yeast. Molecular and General Genetics 261: 977–984Google Scholar
  4. Ambudkar S, Lelong I, Zhang J, Cardarelli C, Gottesman M and Pastan I (1992) Partial Purification and Reconstitution of the Human Multidrug-Resistance Pump: Characterization of the Drug-Stimulatable ATP Hydrolysis. Proceedings of the National Academy of Sciences of United States of America 89: 8472–8476Google Scholar
  5. Ambudkar SV, Dey S, Hrycyna CA, Ramachandra M, Pastan I and Gottesman MM (1999) Biochemical, cellular, and pharmacological aspects of the multidrug transporter. Annual Review of Pharmacology and Toxicology: 39361–39398Google Scholar
  6. Ames GFL, Nikaido K, Groarke J and Petithory J (1989) Reconstitution of periplasmic transport in inside-out membrane vesicles: energization by ATP. Journal of Biological Chemistry 264: 3998–4002Google Scholar
  7. Andrade AC (2000) ABC transporters and multidrug resistance in Aspergillus nidulans. PhD Thesis Wageningen University, Wageningen, the Netherlands, ISBN 90-5808-279-2Google Scholar
  8. Andrade AC, Del Sorbo G, Van Nistelrooy JGM and De Waard MA (2000a) The ABC transporter AtrB from Aspergillus nidulans mediates resistance to all major classes of fungicides and some natural toxic compounds. Microbiology 146: 1987–1997Google Scholar
  9. Andrade AC, Van Nistelrooy JGM, Peery RB, Skatrud PL and De Waard MA (2000b) The role of ABC transporters from Aspergillus nidulans in protection against cytotoxic agents and in antibiotic production. Molecular and General Genetics 263: 966–977Google Scholar
  10. Avendano C and Menendez JC (2002) Inhibitors of multidrug resistance to antitumor agents (MDR). Current Medical Chemistry 9: 159–193Google Scholar
  11. Azzaria M, Schurr E and Gros P (1989) Discrete mutations introduced in the predicted nucleotide-binding sites of the mdr1 gene abolish its ability to confer multidrug resistance. Molecular and Cellular Biology 9: 5289–5297Google Scholar
  12. Bauer BE, Wolfger H and Kuchler K (1999) Inventory and function of yeast ABC proteins: about sex, stress, pleiotropic drug and heavy metal resistance. Biochimica Biophysica Acta, Biomembranes: 217–236Google Scholar
  13. Bissinger PH and Kuchler K (1994) Molecular cloning and expression of the Saccharomyces cerevisiae Sts1 gene product - a yeast ABC transporter conferring mycotoxin resistance. Journal of Biological Chemistry 269: 4180–4186Google Scholar
  14. Bosch I and Croop J (1996) P-glycoprotein multidrug resistance and cancer. Biochimimica Biophysica Acta 1288: F37–F54Google Scholar
  15. Callahan TM, Rose MS, Meade MJ, Ehrenshaft M and Upchurch RG (1999) CFP, the putative cercosporin transporter of Cercospora kikuchii, is required for wild type cercosporin production, resistance, and virulence on soybean. Molecular Plant-Microbe Interactions 12: 901–910Google Scholar
  16. Chang P-K, Yu J, Bhatnagar D and Cleveland TE (1999) The carboxy-terminal portion of the aflatoxin pathway regulatory protein AFLR of Aspergillus parasiticus activates GAL1::lacZ gene expression in Saccharomyces cerevisiae. Applied and Environmental Microbiology 65: 2508–2512Google Scholar
  17. Christensen PU, Davey J and Nielsen O (1997) The Schizosaccharomyces pombe mam1 gene encodes an ABC transporter mediating secretion of M-factor. Molecular and General Genetics 255: 226–236Google Scholar
  18. De Waard MA, Van Nistelrooy JGM, Langeveld CR, Van Kan JAL and Del Sorbo G (1995) Multidrug resistance in filamentous fungi. 11th International Rheinhardsbrunn Symposium: Modern Fungicides and Antifungal Compounds (pp 293–300) Friedrichroda, GermanyGoogle Scholar
  19. De Waard MA (1997) Significance of ABC transporters in fungicide sensitivity and resistance. Pesticide Science 51: 271–275Google Scholar
  20. De Waard MA, Groeneweg H and Van Nistelrooy JGM (1982) Laboratory resistance to fungicides which inhibit ergosterol biosynthesis in Penicillium italicum. Netherlands Journal of Plant Pathology: 99–112Google Scholar
  21. De Waard MA and Van Nistelrooy JGM (1982) Antagonistic and synergistic activities of various chemicals on the toxicity of fenarimol to Aspergillus nidulans. Pesticide Science: 279–286Google Scholar
  22. De Waard MA and Van Nistelrooy JGM (1980) An energy-dependent efflux mechanism for fenarimol in a wild-type strain and fenarimol-resistant mutants of Aspergillus nidulans. Pesticide Biochemistry Psysiology 13: 255–266Google Scholar
  23. De Waard MA and Van Nistelrooy JGM (1984a) Differential accumulation of fenarimol by a wild-type isolate and fenarimol-resistant isolates of Penicillium italicum. Netherlands Journal of Plant Pathology: 143–153Google Scholar
  24. De Waard MA and Van Nistelrooy JGM (1984b) Effects of phthalimide fungicides on the accumulation of fenarimol by Aspergillus nidulans. Pesticide Science: 56–62Google Scholar
  25. De Waard MA and Van Nistelrooy JGM (1988) Accumulation of SBI fungicides in wild-type and fenarimol-resistant isolates of Penicillium italicum. Pesticide Science 22: 371–382Google Scholar
  26. Decottignies A and Goffeau A (1997) Complete inventory of the yeast ABC proteins. Nature Genetics 15: 137–145Google Scholar
  27. Dekker J (1981) Impact of fungicide resistance of disease control toxicity. British Crop Protection Conference: pests and diseases (pp 857–863) Brighton, EnglandGoogle Scholar
  28. Del Sorbo G, Andrade AC, Van Nistelrooy JG, Van Kan JA, Balzi E and De Waard MA (1997) Multidrug resistance in Aspergillus nidulans involves novel ATP-binding cassette transporters. Molecular and General Genetics 254: 417–426Google Scholar
  29. Del Sorbo G, Schoonbeek H and De Waard MA (2000) Fungal transporters involved in efflux of natural toxic compounds and fungicides. Fungal Genetics and Biology 30: 1–15Google Scholar
  30. Denning DW, Venkateswarlu K, Oakley KL, Anderson MJ, Manning NJ, Stevens DA, Warnock DW and Kelly SL (1997) Itraconazole resistance in Aspergillus fumigatus. Antimicrobial Agents and Chemotherapy 41: 1364–1368Google Scholar
  31. Desjardins AE, Proctor RH, Bai G, McCormick SP, Shaner G, Buechley G and Hohn TM (1996) Reduced virulence of trichothecene-nonproducing mutants of Gibberella zeae in wheat field tests. Molecular Plant-Microbe Interactions 9: 775–781Google Scholar
  32. Do Nascimiento AM, Terenzi MF, Goldman MMH and Goldman GH (1999) A novel ATP-binding cassette transporter involved in multidrug resistance in the filamentous fungus Aspergillus nidulans. Fungal Genetics Newsletter 46: 44Google Scholar
  33. Fleissner A, Sopalla C and Weltring K (2002) An ATP-binding cassette multidrug-resistance transporter is necessary for tolerance of Gibberella pulicaris to phytoalexins and virulence on potato tubers. Molecular Plant-Microbe Interactions 15: 102–108Google Scholar
  34. Fling ME, Kopf J, Tamarkin A, Gorman JA, Smith HA and Koltin Y (1991) Analysis of a Candida albicans gene that encodes a novel mechanism for resistance to benomyl and methotrexate. Molecular and General Genetics 227: 318–329Google Scholar
  35. Franz R, Kelly SL, Lamb DC, Kelly DE, Ruhnke M and Morschhauser J (1998) Multiple molecular mechanisms contribute to a stepwise development of fluconazole resistance in clinical Candida albicans strains. Antimicrobial Agents and Chemotherapy 42: 3065–3072Google Scholar
  36. Gottesman MM and Pastan I (1988) The multidrug transporter, a double-edged sword. Journal of Biological Chemistry 263: 12163–12166Google Scholar
  37. Gottesman MM and Pastan I (1993) Biochemistry of multidrug resistance mediated by the multidrug transporter. Annual Review of Biochemistry 62: 385–427Google Scholar
  38. Greenberger LM (1993) Major photoaffinity drug labeling sites for iodoaryl azidoprazosin in P-glycoprotein are within, or immediately C-terminal to, transmembrane domains 6 and 12. Journal of Biological Chemistry 268: 11417–11425Google Scholar
  39. Griffith JK, Baker ME, Rouch DA, Page MG, Skurray RA, Paulsen IT, Chater KF, Baldwin SA and Henderson PJ (1992) Membrane transport proteins: implications of sequence comparisons. Current Opinion in Cell Biology 4: 684–695Google Scholar
  40. Hamamoto H, Hasegawa K, Nakaune R, Lee YJ, Makizumi Y, Akutsu K and Hibi T (2000) Tandem repeat of a transcriptional enhancer upstream of the sterol 14alpha-demethylase gene (CYP51) in Penicillium digitatum. Applied and Environmental Microbiology 66: 3421–3426Google Scholar
  41. Hayashi K, Schoonbeek H and De Waard MA (2002) Bcmfs1, a novel ABC transporter from Botrytis cinerea, provides tolerance to the natural toxic compounds camptothecin and to fungicides (submitted)Google Scholar
  42. Hayashi K, Schoonbeek H, Sugiura H and De Waard MA (2001) Multidrug resistance in Botrytis cinerea associated with decreased accumulation of the azole fungicide oxpoconazole and increased transcription of the ABC transporter gene BcatrD. Pesticide Biochemistry and Physiology 70: 168–179Google Scholar
  43. Henderson P (1993) The 12-transmembrane helix transporters. Current Opinion in Cell Biology 5: 708–721Google Scholar
  44. Higgins CF and Gottesman M (1992) Is the multidrug transporter a flippase? Trends in Biochemical Sciences 17: 18–21Google Scholar
  45. Higgins CF (1992) ABC transporters: from microorganisms to man. Annual Review of Cell Biology 8: 67–113Google Scholar
  46. Ishikawa T (1992) The ATP-dependent glutathione S-conjugate export-pump. Trends in Biochemical Sciences 17: 463–468Google Scholar
  47. Ishikawa T, Li ZS, Lu YP and Rea PA (1997) The GS-X pump in plant, yeast, and animal cells: structure, function, and gene expression. Bioscience Reports 17: 189–207Google Scholar
  48. Juliano RL and Ling V (1976) A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. Biochimica Biophysica Acta 455: 152–162Google Scholar
  49. Kema GHJ, Yu D, Rijkenberg FHJ, Shaw MW and Baayen RP (1996) Histology of the pathogenesis of Mycosphaerella graminicola in wheat. Phytopathology 86: 777–786Google Scholar
  50. Kilty JE and Amara SG (1992) Families of twelve transmembrane domain transporters. Current Opinion in Biotechnology 3: 675–682Google Scholar
  51. Kuchler K, Sterne RE and Thorner J (1989) Saccharomyces cerevisiae STE6 gene product: A novel pathway for protein export in eukaryotic cells. The EMBO Journal 8: 3973–3984Google Scholar
  52. Loe DW, Stewart RK, Massey TE, Deeley RG and Cole SP (1997) ATP-dependent transport of aflatoxin B1 and its glutathione conjugates by the product of the multidrug resistance protein (MRP) gene. Molecular Pharmacology 51: 1034–1041Google Scholar
  53. Loo TW and Clarke DM (1994) Reconstitution of drug-stimulated ATPase activity following co-expression of each half of human P-glycoprotein as separate polypeptides. Journal of Biological Chemistry 269: 7750–7755Google Scholar
  54. Loo TW and Clarke DM (1995) Covalent modification of human P-glycoprotein mutants containing a single cysteine in either nucleotide-binding fold abolishes drug-stimulated ATPase activity. Journal of Biological Chemistry 270: 22957–22961Google Scholar
  55. Marger MD and Saier MHJ (1993) A major superfamily of transmembrane facilitators that catalyse uniport, symport and antiport. Trends in Biochemical Sciences 18: 13–20Google Scholar
  56. Muhitch MJ, McCormick SP, Alexander NJ and Hohn TM (2000) Transgenic expression of the TRI101 or PDR5 gene increases resistance of tobacco to the phytotoxic effects of the trichothecene 4,15-diacetoxyscirpenol. Plant Science 157: 201–207Google Scholar
  57. Nakajima M, Suzuki J, Hosaka T, Hibi T and Akutsu K (2001) Functional analysis of anATP-binding cassette transporter gene in Botrytis cinerea by gene disruption. Journal of General Plant Pathology 67: 212–214Google Scholar
  58. Nakaune R, Adachi K, Nawata O, Tomiyama M, Akutsu K and Hibi T (1998) A novel ATP-binding cassette transporter involved in multidrug resistance in the phytopathogenic fungus Penicillium digitatum. Applied and Environmental Microbiology 64: 3983–3988Google Scholar
  59. Nakaune R, Hamamoto H, Imada J, Akutsu K and Hibi T (2002) A novel ABC transporter gene, PMR5, is involved in multidrug resistance in the phytopathogenic fungus Penicillium digitatum. Molecular Genetics and Genomics: Published online: 22 FebruaryGoogle Scholar
  60. Nelissen B, De Wachter R and Goffeau A (1997) Classification of all putative permeases and other membrane plurispanners of the major facilitator superfamily encoded by the complete genome of Saccharomyces cerevisiae. FEMS Microbiology Reviews 21: 113–134Google Scholar
  61. Pao SS, Paulsen IT and Saier MH, Jr. (1998) Major facilitator superfamily. Microbiology and Molecular Biology Reviews 62: 1–34Google Scholar
  62. Paulsen IT, Brown MH and Skurray RA (1996) Proton-dependent multidrug efflux systems. Microbiology Reviews 60: 575–608Google Scholar
  63. Paulsen IT and Skurray RA (1993) Topology, structure and evolution of two families of proteins involved in antibiotic and antiseptic resistance in eukaryotes and prokaryotes - an analysis. Gene 132: 155Google Scholar
  64. Perea S, Lopez-Ribot JL, Kirkpatrick WR, McAtee RK, Santillan RA, Martinez M, Calabrese D, Sanglard D and Patterson TF (2001) Prevalence of molecular mechanisms of resistance to azole antifungal agents in Candida albicans strains displaying high-level fluconazole resistance isolated from human immunodeficiency virus-infected patients. Antimicrobial Agents and Chemotherapy 45: 2676–2684Google Scholar
  65. Pitkin JW, Panaccione DG and Walton JD (1996) A putative cyclic peptide efflux pump encoded by the TOXA gene of the plantpathogenic fungus Cochliobolus carbonum. Microbiology 142: 1557–1565Google Scholar
  66. Raviv Y, Pollard H, Bruggemann E, Pastan I and Gottesman M (1990) Photosensitized labeling of a functional multidrug transporter in living drug-resistant tumor cells. Journal of Biological Chemistry 265: 3975–3980Google Scholar
  67. Roepe P (1994) Indirect mechanism of drug transport by P-glycoprotein. Trends in Pharmacological Sciences 15: 445–446Google Scholar
  68. Rosenberg MF, Callaghan R, Ford RC and Higgins CF (1997) Structure of the multidrug resistance P-glycoprotein to 2.5 nm resolution determined by electron microscopy and image analysis. Journal of Biological Chemistry 272: 10685–10694Google Scholar
  69. Rubin RA, Levy SB, Heinrikson RL and Kezdy FJ (1990) Gene duplication in the evolution of the two complementing domains of gram-negative bacterial tetracycline efflux proteins. Gene 87: 7–13Google Scholar
  70. Safa AR, Stern RK, Choi K, Agresti M, Tamai I, Mehta ND and Roninson IB (1990) Molecular basis of preferential resistance to colchicine in multidrug-resistant human cells conferred by Gly-185 fwdarw Val-185 substitution in P-glycoprotein. Proceedings of the National Academy of Sciences of the United States of America 87: 7225–7229Google Scholar
  71. Saier MH, Jr., Paulsen IT, Sliwinski MK, Pao SS, Skurray RA and Nikaido H (1998) Evolutionary origins of multidrug and drug-specific efflux pumps in bacteria. The FASEB Journal 12: 265–274Google Scholar
  72. Sanglard D, Ischer F, Monod M and Bille J (1997) Cloning of Candida albicans genes conferring resistance to azole antifungal agents: Characterization of CDR2, a new multidrug ABC transporter gene. Microbiology 143: 405–416Google Scholar
  73. Sanglard D, Kuchler K, Ischer F, Pagani JL, Monod M and Bille J (1995) Mechanisms of resistance to azole antifungal agents in Candida albicans isolates from AIDS patients involve specific multidrug transporters. Antimicrobial Agents and Chemotherapy 39: 2378–2386Google Scholar
  74. Schnabel G and Jones LA (2001) Fungicide resistance genes in Venturia inaequalis. Modern fungicides and antifungal compounds. 13th International Reinhardsbrunn Symposium: Modern Fungicides and Antifungal Compounds, Friedrichroda, GermanyGoogle Scholar
  75. Schoonbeek H, Del Sorbo G and De Waard MA (2001) The ABC transporter BcatrB affects the sensitivity of Botrytis cinerea to the phytoalexin resveratrol and the fungicide fenpiclonil. Molecular Plant-Microbe Interactions 14: 562–571Google Scholar
  76. Slaven JW, Anderson MJ, Sanglard D, Dixon GK, Bille J, Roberts IA and Denning DW (1999) Fungal Genetics Newsletter 46: 64Google Scholar
  77. St Georgiev V (2000) Membrane transporters and antifungal drug resistance. Current Drug Targets 1: 261–284Google Scholar
  78. Stergiopoulos I, Gielkens MMC, Goodall SD, Venema K and De Waard MA (2002a) ABC transporter genes from the wheat pathogen Mycosphaerella graminicola. Gene 289: 141–149Google Scholar
  79. Stergiopoulos I, Zwiers L-H and De Waard MA (2002b) ABC transporters involved in pathogenesis of Mycosphaerella graminicola. 6th European Conference of Fungal Genetics (p 250) Pisa, ItalyGoogle Scholar
  80. Taglicht D and Michaelis S (1998) Saccharomyces cerevisiae ABC proteins and their relevance to human health and disease. Methods in Enzymology 292: 130–162Google Scholar
  81. Taylor JL and Condie J (1999) Characterisation of ABC transporters from the fungal phytopathogen Leptosphaeria maculans. 9th International Congress on Molecular Plant-Microbe Interactions (p L71) Amsterdam, the NetherlandsGoogle Scholar
  82. Theodoulou FL (2000) Plant ABC transporters. Biochimica Biophysica Acta, Biomembranes 1465: 79–103Google Scholar
  83. Tobin MB, Peery RB and Skatrud PL (1997) Genes encoding multiple drug resistance-like proteins in Aspergillus fumigatus and Aspergillus flavus. Gene 200: 11–23Google Scholar
  84. Tusnady GE, Bakos E, Varadi A and Sarkadi B (1997) Membrane topology distinguishes a subfamily of theATP-binding cassette (ABC) transporters. FEBS Letters 402: 1–3Google Scholar
  85. Upchurch RG, Rose MS and Eweida M (2001) Over-expression of the cercosporin facilitator protein, CFP, in Cercospora kikuchii up-regulates production and secretion of cercosporin. FEMS Microbiology Letters 204: 89–93Google Scholar
  86. Urban M, Bhargava T and Hamer JE (1999) An ATP-driven efflux pump is a novel pathogenicity factor in rice blast disease. The EMBO Journal 18: 512–521Google Scholar
  87. Vanden Bossche H, Dromer F, Improvisi I, Lozano-Chiu M, Rex JH and Sanglard D (1998) Antifungal drug resistance in pathogenic fungi. Medical Mycology 36: 119–128Google Scholar
  88. Vermeulen T, Schoonbeek H and De Waard MA (2001) The ABC transporter BcatrB from Botrytis cinerea is a determinant of the phenylpyrrole fungicide fludioxonil. Pest Management Science 57: 393–402Google Scholar
  89. Voss T, Schulte J and Tudzynski B (2001) A new MFS-transporter gene next to the gibberellin biosynthesis gene cluster of Gibberella fujikuroi is not involved in gibberellin secretion. Current Genetics 39: 377–383Google Scholar
  90. Wadkins RM RP (1997) Biophysical aspects of P-glycoproteinmediated multidrug resistance. International Review of Cytology 171: 121–165Google Scholar
  91. Walker JE, Saraste M, Runswick MJ and Gay NJ (1982) Distantly related sequences in the alpha-and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. The EMBO Journal 1: 945–951Google Scholar
  92. White T (1997) Increased mRNA levels of ERG16, CDR, and MDR1 correlate with increases in azole resistance in Candida albicans isolates from a patient infected with human immunodeficiency virus. Antimicrobial Agents and Chemotherapy 41: 1482–1487Google Scholar
  93. Wolfger H, MamnunYM and Kuchler K (2001) Fungal ABC proteins: pleiotropic drug resistance, stress response and cellular detoxification. Research in Microbiology 152: 375–389Google Scholar
  94. Yoder OC and Turgeon BG (2001) Fungal genomics and pathogenicity. Current Opinion in Plant Biology 4: 315–321Google Scholar
  95. Zhang XP, Collins KI and Greenberger LM (1995) Functional evidence that transmembrane 12 and the loop between transmembrane 11 and 12 form part of the drug-binding domain in P-glycoprotein encoded by MDR1. Journal of Biological Chemistry: 5441–5448Google Scholar
  96. Zwiers L-H (2002) ABC transporters of the wheat pathogen Mycosphaerella graminicola. PhD Thesis Wageningen University, Wageningen, the Netherlands, ISBN 90-5808-613-5Google Scholar
  97. Zwiers LH and De Waard MA (2000) Characterization of the ABC transporter genes MgAtr1 and MgAtr2 from the wheat pathogen Mycosphaerella graminicola. Fungal Genetics and Biology 30: 115–125Google Scholar

Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Ioannis Stergiopoulos
    • 1
  • Lute-Harm Zwiers
    • 1
  • Maarten A. De Waard
    • 1
  1. 1.Laboratory of Phytopathology, Department of Plant SciencesWageningen UniversityWageningenThe Netherlands

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