Applied Microbiology and Biotechnology

, Volume 67, Issue 6, pp 807–815 | Cite as

Examination of fungal stress response genes using Saccharomyces cerevisiae as a model system: targeting genes affecting aflatoxin biosynthesis by Aspergillus flavus Link

  • Jong H. Kim
  • Bruce C. Campbell
  • Jiujiang Yu
  • Noreen Mahoney
  • Kathleen L. Chan
  • Russell J. Molyneux
  • Deepak Bhatnagar
  • Thomas E. Cleveland
Applied Genetics and Molecular Biotechnology

Abstract

Saccharomyces cerevisiae served as a model fungal system to examine functional genomics of oxidative stress responses and reactions to test antioxidant compounds. Twenty-two strains of S. cerevisiae, including a broad spectrum of singular gene deletion mutants, were exposed to hydrogen peroxide (H2O2) to examine phenotypic response to oxidative stress. Responses of particular mutants treated with gallic, tannic or caffeic acids, or methyl gallate, during H2O2 exposure, indicated that these compounds alleviated oxidative stress. These compounds are also potent inhibitors of aflatoxin biosynthesis in Aspergillus flavus. To gain further insights into a potential link between oxidative stress and aflatoxin biosynthesis, 43 orthologs of S. cerevisiae genes involved in gene regulation, signal transduction (e.g., SHO1, HOG1, etc.) and antioxidation (e.g., CTT1, CTA1, etc.) were identified in an A. flavus expressed sequence tag library. A successful exemplary functional complementation of an antioxidative stress gene from A. flavus, mitochondrial superoxide dismutase (sodA), in a sod2Δ yeast mutant further supported the potential of S. cerevisiae deletion mutants to serve as a model system to study A. flavus. Use of this system to further examine functional genomics of oxidative stress in aflatoxigenesis and reduction of aflatoxin biosynthesis by antioxidants is discussed.

References

  1. Bhatnagar D, Yu J, Ehrlich KC (2002) Toxins of filamentous fungi. Chem Immunol 81:167–206PubMedGoogle Scholar
  2. Bok JW, Keller NP (2004) LaeA, a regulator of secondary metabolism in Aspergillus spp. Eukaryot Cell 3:527–535PubMedGoogle Scholar
  3. Bolwell GP (1999) Role of active oxygen species and NO in plant defense responses. Curr Opin Plant Biol 2:287–294PubMedGoogle Scholar
  4. Campbell BC, Molyneux RJ, Schatzki TF (2003) Current research on reducing pre- and post-harvest aflatoxin contamination of U. S. almond, pistachio and walnut. In: Abbas H (ed) Aflatoxin and food safety. Part I. J Toxicol-Toxin Rev 22:225–266Google Scholar
  5. Cary JF, Harris PY, Molyneux RJ, Mahoney NE (2003) Inhibition of aflatoxin biosynthesis by gallic acid. In: Proceedings of the 3rd Fungal Genomics, 4th Fumonisin, and 16th Aflatoxin Elimination Workshop. 13–15 October, 2003, Savannah, GA., p 51Google Scholar
  6. Estruch F (2000) Stress-controlled transcription factors, stress-induced genes and stress tolerance in budding yeast. FEMS Microbiol Rev 24:469–486CrossRefPubMedGoogle Scholar
  7. Fernandes L, Rodrigues-Pousada C, Struhl K (1997) Yap, a novel family of eight bZIP proteins in Saccharomyces cerevisiae with distinct biological functions. Mol Cell Biol 17:6982–6993PubMedGoogle Scholar
  8. Grant CM, MacIver FH, Dawes IW (1997) Mitochondrial function is required for resistance to oxidative stress in the yeast Saccharomyces cerevisiae. FEBS Lett 410:219–222PubMedGoogle Scholar
  9. Jayashree T, Subramanyam C (1999) Antiaflatoxigenic activity of eugenol is due to inhibition of lipid peroxidation. Lett Appl Microbiol 28:179–183PubMedGoogle Scholar
  10. Jayashree T, Subramanyam C (2000) Oxidative stress as a prerequisite for aflatoxin production by Aspergillus parasiticus. Free Radic Biol Med 29:981–985PubMedGoogle Scholar
  11. Kozak M (1987) An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res 15:8125–8148PubMedGoogle Scholar
  12. Lee J, Godon C, Lagniel G, Spector D, Garin J, Labarre J, Toledano MB (1999) Yap1 and Skn7 control two specialized oxidative stress response regulons in yeast. J Biol Chem 274:16040–16046PubMedGoogle Scholar
  13. Levine A, Tenhaken R, Dixon R, Lamb C (1994) H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79:583–593CrossRefPubMedGoogle Scholar
  14. Mahoney N, Molyneux RJ (2004) Phytochemical inhibition of aflatoxigenicity in Aspergillus flavus by constituents of walnut (Juglans regia). J Agric Food Chem 52:1882–1889PubMedGoogle Scholar
  15. Marchler-Bauer A, Anderson JB, DeWeese-Scott C, Fedorova ND, Geer LY, He S, Hurwitz DI, Jackson JD, Jacobs AR, Lanczycki CJ, Liebert CA, Liu C, Madej T, Marchler GH, Mazumder R, Nikolskaya AN, Panchenko AR, Rao BS, Shoemaker BA, Simonyan V, Song JS, Thiessen PA, Vasudevan S, Wang Y, Yamashita RA, Yin JJ, Bryant SH (2003) CDD: a curated Entrez database of conserved domain alignments. Nucleic Acids Res 31:383–387CrossRefPubMedGoogle Scholar
  16. Skory CD, Chang PK, Cary J, Linz JE (1992) Isolation and characterization of a gene from Aspergillus parasiticus associated with the conversion of versicolorin A to sterigmatocystin in aflatoxin biosynthesis. Appl Environ Microbiol 58: 3527–3537Google Scholar
  17. Sroka Z, Cisowski W (2003) Hydrogen peroxide scavenging, antioxidant and anti-radical activity of some phenolic acids. Food Chem Toxicol 41:753–758PubMedGoogle Scholar
  18. Sweeney MJ, Dobson AD (1999) Molecular biology of mycotoxin biosynthesis. FEMS Microbiol Lett 175:149–163PubMedGoogle Scholar
  19. Toone WM, Jones N (1998) Stress-activated signaling pathways in yeast. Genes Cells 3:485–498PubMedGoogle Scholar
  20. Tucker CL, Fields S (2004) Quantitative genome-wide analysis of yeast deletion strain sensitivities to oxidative and chemical stress. Comp Funct Genomics 5:216–224Google Scholar
  21. Vogel HJ (1956) A convenient growth medium for Neurospora (medium N). Microb Genet Bull 13:42–44Google Scholar
  22. Wang W, Vinocur B, Altman A (2003) Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218:1–14CrossRefPubMedGoogle Scholar
  23. Winzeler EA, Shoemaker DD, Astromoff A et al (1999) Functional characterization of the S. cerevisiae genome by gene deletion and parallel analysis. Science 285:901–906CrossRefPubMedGoogle Scholar
  24. Ying W, Swanson RA (2000) The poly(ADP-ribose) glycohydrolase inhibitor gallotannin blocks oxidative astrocyte death. Neuroreport 11:1385–1388PubMedGoogle Scholar
  25. Ying W, Sevigny MB, Chen Y, Swanson RA (2001) Poly(ADP-ribose) glycohydrolase mediates oxidative and excitotoxic neuronal death. Proc Natl Acad Sci USA 98:12227–12232Google Scholar
  26. Yu J, Chang P-K, Ehrlich KC, Cary JW, Bhatnagar D, Cleveland TE, Payne GA, Linz JE, Woloshuk CP, Bennett JW (2004a) Clustered pathway genes in aflatoxin biosynthesis. Appl Environ Microbiol 70:1253–1262PubMedGoogle Scholar
  27. Yu J, Whitelaw CA, Nierman WC, Bhatnagar D, Cleveland TE (2004b) Aspergillus flavus expressed sequence tags for identification of genes with putative roles in aflatoxin contamination of crops. FEMS Microbiol Lett 237:333–340PubMedGoogle Scholar
  28. Zelko IN, Mariani TJ, Folz RJ (2002) Superoxide dismutase multigene family: a comparison of the CuZn-SOD (SOD1), Mn-SOD (SOD2), and EC-SOD (SOD3) gene structures, evolution, and expression. Free Radic Biol Med 33:337–349PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Jong H. Kim
    • 1
  • Bruce C. Campbell
    • 1
  • Jiujiang Yu
    • 2
  • Noreen Mahoney
    • 1
  • Kathleen L. Chan
    • 1
  • Russell J. Molyneux
    • 1
  • Deepak Bhatnagar
    • 2
  • Thomas E. Cleveland
    • 2
  1. 1.Plant Mycotoxin Research UnitWestern Regional Research Center, USDA-ARSAlbanyUSA
  2. 2.Food and Feed Safety Research UnitSouthern Regional Research Center, USDA-ARSNew OrleansUSA

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