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A Polyketide Synthase Gene Required for Biosynthesis of the Aflatoxin-like Toxin, Dothistromin

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Abstract

Dothistromin is a polyketide toxin, produced by a fungal forest pathogen, with structural similarity to the aflatoxin precursor versicolorin B. Biochemical and genetic studies suggested that there are common steps in the biosynthetic pathways for these metabolites and showed similarities between some of the genes. A polyketide synthase gene (pksA) was isolated from dothistromin-producing Dothistroma septosporum by hybridization with an aflatoxin ortholog from Aspergillus parasiticus. Inactivation of this gene in D. septosporum resulted in mutants that could not produce dothistromin but that could convert exogenous aflatoxin precursors, including norsolorinic acid, into dothistromin. The mutants also had reduced asexual sporulation compared to the wild type. So far four other genes are known to be clustered immediately alongside pksA. Three of these (cypA, moxA, avfA) are predicted to be orthologs of aflatoxin biosynthetic genes. The other gene (epoA), located between avfA and moxA, is predicted to encode an epoxide hydrolase, for which there is no homolog in either the aflatoxin or sterigmatocystin gene clusters. The pksA gene is located on a small chromosome of ~1.3 Mb in size, along with the dothistromin ketoreductase (dotA) gene.

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References

  1. RE Bradshaw (2004) ArticleTitle Dothistroma (red-band) needle blight of pines and the dothistromin toxin: a review For Pathol 34 163–185

    Google Scholar 

  2. L Shain RA Franich (1981) ArticleTitleInduction of Dothistroma blight symptoms with dothistromin Physiol Plant Pathol 19 49–55 Occurrence Handle1:CAS:528:DyaL3MXlsVOgt78%3D

    CAS  Google Scholar 

  3. JW Bennett MA Klich (2003) ArticleTitleMycotoxins Clin Microbiol Rev 16 497–516 Occurrence Handle10.1128/CMR.16.3.497-516.2003 Occurrence Handle1:CAS:528:DC%2BD3sXmsV2qtL4%3D Occurrence Handle12857779

    Article  CAS  PubMed  Google Scholar 

  4. RA Squire (1989) ArticleTitleRanking animal carcinogens: a proposed regulatory approach Science 214 887–891

    Google Scholar 

  5. TE Cleveland PF Dowd AE Desjardins D Bhatnagar PJ Cotty (2003) ArticleTitleUnited States Department of Agriculture – Agricultural Research Service research on pre-harvest prevention of mycotoxins and mycotoxigenic fungi in US crops Pest Manag Sci 59 629–642 Occurrence Handle10.1002/ps.724 Occurrence Handle1:CAS:528:DC%2BD3sXkslektr4%3D Occurrence Handle12846313

    Article  CAS  PubMed  Google Scholar 

  6. J Yu D Bhatnagar TE Cleveland (2004) ArticleTitleCompleted sequence of aflatoxin pathway gene cluster in Aspergillus parasiticus FEBS Lett 564 126–130 Occurrence Handle10.1016/S0014-5793(04)00327-8 Occurrence Handle1:CAS:528:DC%2BD2cXjt1Cgurk%3D Occurrence Handle15094053

    Article  CAS  PubMed  Google Scholar 

  7. DW Brown J-H Yu HS Kelkar M Fernandes TC Nesbitt NP Keller TH Adams TJ Leonard (1996) ArticleTitleTwenty-five co-regulated transcripts define a sterigmatocystin gene cluster in Aspergillus nidulans Proc Natl Acad Sci USA 93 1418–1422 Occurrence Handle10.1073/pnas.93.4.1418 Occurrence Handle1:CAS:528:DyaK28Xht1ens7w%3D Occurrence Handle8643646

    Article  CAS  PubMed  Google Scholar 

  8. NP Keller TM Hohn (1997) ArticleTitleMetabolic pathway gene clusters in filamentous fungi Fungal Genet Biol 21 17–29 Occurrence Handle10.1006/fgbi.1997.0970 Occurrence Handle1:CAS:528:DyaK2sXitlKlsL0%3D Occurrence Handle9073477

    Article  CAS  PubMed  Google Scholar 

  9. GJ Shaw M Chick R Hodges (1978) ArticleTitleA 13C-NMR study of the biosynthesis of the anthraquinone dothistromin by Dothistroma pini Phytochemistry 17 1743–1745 Occurrence Handle10.1016/S0031-9422(00)88685-9 Occurrence Handle1:CAS:528:DyaE1MXhvVaru7o%3D

    Article  CAS  Google Scholar 

  10. RE Bradshaw D Bhatnagar RJ Ganley CJ Gillman BJ Monahan JM Seconi (2002) ArticleTitle Dothistroma pini, a forest pathogen, contains homologs of aflatoxin biosynthetic pathway genes Appl Environ Microbiol 68 2885–2892 Occurrence Handle10.1128/AEM.68.6.2885-2892.2002 Occurrence Handle1:CAS:528:DC%2BD38XksVWqsbY%3D Occurrence Handle12039746

    Article  CAS  PubMed  Google Scholar 

  11. I Barnes PW Crous BD Wingfield MJ Wingfield (2004) ArticleTitleMultigene phylogenies reveal that red band needle blight of Pinus is caused by two distinct species of Dothistroma, D. septosporum and D. pini Stud Mycol 50 551–565

    Google Scholar 

  12. S Kroken NL Glass JW Taylor OC Yoder BG Turgeon (2003) ArticleTitlePhylogenomic analysis of type I polyketide synthase genes in pathogenic and saprobic ascomycetes Proc Natl Acad Sci USA 100 15,670–15,675 Occurrence Handle10.1073/pnas.2532165100 Occurrence Handle1:CAS:528:DC%2BD2cXhtVCgtw%3D%3D

    Article  CAS  Google Scholar 

  13. P-K Chang JW Cary J Yu D Bhatnagar TE Cleveland (1995) ArticleTitleThe Aspergillus parasiticus polyketide synthase gene pksA, a homolog of Aspergillus nidulans wA, is required for aflatoxin B1 biosynthesis Mol Gen Genet 248 270–277 Occurrence Handle10.1007/BF02191593 Occurrence Handle1:CAS:528:DyaK2MXot1Gmtbk%3D Occurrence Handle7565588

    Article  CAS  PubMed  Google Scholar 

  14. RE Bradshaw RJ Ganley WT Jones PS Dyer (2000) ArticleTitleHigh levels of dothistromin toxin produced by the forest pathogen Dothistroma pini Mycol Res 104 325–332 Occurrence Handle10.1017/S0953756299001367 Occurrence Handle1:CAS:528:DC%2BD3cXjtVyms70%3D

    Article  CAS  Google Scholar 

  15. RA Franich (1981) ArticleTitleDetermination of dothistromin by quantitative reversed-phase thin-layer chromatography J Chromatogr 209 117–120 Occurrence Handle10.1016/S0021-9673(00)80435-5 Occurrence Handle1:CAS:528:DyaL3MXitVCks7Y%3D

    Article  CAS  Google Scholar 

  16. SF Altschul TL Madden AA Schaffer J Zhang Z Zhang W Miller DJ Lipman (1997) ArticleTitleGapped BLAST, PSI-BLAST: a new generation of protein database search programs Nucleic Acids Res 25 3389–3402 Occurrence Handle10.1093/nar/25.17.3389 Occurrence Handle1:CAS:528:DyaK2sXlvFyhu7w%3D Occurrence Handle9254694

    Article  CAS  PubMed  Google Scholar 

  17. JJ Doyle JL Doyle (1987) ArticleTitleA rapid DNA isolation procedure for small quantities of fresh leaf tissue Phytochem Bull 19 11–15

    Google Scholar 

  18. RE Bradshaw A Bidlake N Forester DB Scott (1997) ArticleTitleTransformation of the fungal forest pathogen Dothistroma pini to hygromycin resistance Mycol Res 101 1247–1250 Occurrence Handle10.1017/S0953756297003870 Occurrence Handle1:CAS:528:DyaK2sXnt1Khsbg%3D

    Article  CAS  Google Scholar 

  19. J Sambrook EF Fritsch T Maniatis (1989) Molecular Cloning: A Laboratory Manual Cold Spring Harbor Laboratory Press New York

    Google Scholar 

  20. WT Jones D Harvey SD Jones S Fielder P Debnam PHS Reynolds (1993) ArticleTitleCompetitive ELISA employing monoclonal antibodies specific for dothistromin Food Agric Immunol 5 187–197 Occurrence Handle1:CAS:528:DyaK2cXkt1yltbs%3D Occurrence Handle10.1080/09540109309354798

    Article  CAS  Google Scholar 

  21. C Young Y Itoh R Johnson I Garthwaite CO Miles SC Munday-Finch DB Scott (1998) ArticleTitlePaxilline-negative mutants of Penicillium paxilli generated by heterologous and homologous plasmid integration Curr Genet 33 368–377 Occurrence Handle10.1007/s002940050349 Occurrence Handle1:CAS:528:DyaK1cXjsVemsrg%3D Occurrence Handle9618588

    Article  CAS  PubMed  Google Scholar 

  22. DW Udwary M Merski CA Townsend (2002) ArticleTitleA method for prediction of the locations of linker regions within large multifunctional proteins, and application to a type I polyketide synthase J Mol Biol 323 585–598 Occurrence Handle10.1016/S0022-2836(02)00972-5 Occurrence Handle1:CAS:528:DC%2BD38XnslGrtLg%3D Occurrence Handle12381311

    Article  CAS  PubMed  Google Scholar 

  23. J Yu P-K Chang KC Ehrlich JW Cary D Bhatnagar TE Cleveland PA Payne JE Linz CP Woloshuk JW Bennett (2004) ArticleTitleClustered pathway genes in aflatoxin biosynthesis Appl Environ Microbiol 70 1253–1262 Occurrence Handle10.1128/AEM.70.3.1253-1262.2004 Occurrence Handle1:CAS:528:DC%2BD2cXisVKjtLg%3D Occurrence Handle15006741

    Article  CAS  PubMed  Google Scholar 

  24. CMH Watanabe CA Townsend (2002) ArticleTitleInitial characterization of a type I fatty acid synthase and polyketide synthase multienzyme complex NorS in the biosynthesis of aflatoxin B1 Chem Biol 9 981–988 Occurrence Handle10.1016/S1074-5521(02)00213-2 Occurrence Handle1:CAS:528:DC%2BD38Xnt1Ggur8%3D Occurrence Handle12323372

    Article  CAS  PubMed  Google Scholar 

  25. ME Mayorga WE Timberlake (1992) ArticleTitleThe developmentally regulated Aspergillus nidulans wA gene encodes a polypeptide homologous to polyketide and fatty acid synthases Mol Gen Genet 235 205–212 Occurrence Handle10.1007/BF00279362 Occurrence Handle1:CAS:528:DyaK3sXlsFejtrs%3D Occurrence Handle1465094

    Article  CAS  PubMed  Google Scholar 

  26. I Fujii A Watanabe U Sankawa Y Ebizuka (2001) ArticleTitleIdentification of Claisen cyclase domain in fungal polyketide synthase WA, a naphthopyrone synthase of Aspergillus nidulans Chem Biol 8 189–197 Occurrence Handle10.1016/S1074-5521(00)90068-1 Occurrence Handle1:CAS:528:DC%2BD3MXjtV2ku78%3D Occurrence Handle11251292

    Article  CAS  PubMed  Google Scholar 

  27. J Yu CP Woloshuk D Bhatnagar TE Cleveland (2000) ArticleTitleCloning and characterization of avfA and omtB genes involved in aflatoxin biosynthesis in three Aspergillus species Gene 248 157–167 Occurrence Handle10.1016/S0378-1119(00)00126-8 Occurrence Handle1:CAS:528:DC%2BD3cXjtVWqu7Y%3D Occurrence Handle10806361

    Article  CAS  PubMed  Google Scholar 

  28. Y Wen H Hatabayashi H Arai K Kitamoto K Yabe (2005) ArticleTitleFunction of the cypX and moxY genes in aflatoxin biosynthesis in Aspergillus parasiticus Appl Environ Microbiol 71 3192–3198 Occurrence Handle10.1128/AEM.71.6.3192-3198.2005 Occurrence Handle1:CAS:528:DC%2BD2MXlt1OkurY%3D Occurrence Handle15933021

    Article  CAS  PubMed  Google Scholar 

  29. M Arand H Hemmer H Durk J Baratti A Archelas R Furtoss F Oesch (1999) ArticleTitleCloning and molecular characterization of a soluble epoxide hydrolase from Aspergillus niger that is related to mammalian microsomal epoxide hydrolase Biochem J 344 273–280 Occurrence Handle10.1042/0264-6021:3440273 Occurrence Handle1:CAS:528:DyaK1MXns1yjtLc%3D Occurrence Handle10548561

    Article  CAS  PubMed  Google Scholar 

  30. EJ Kelly KE Erickson C Sengstag DL Eaton (2002) ArticleTitleExpression of human microsomal epoxide hydrolase in Saccharomyces cerevisiae reveals a functional role in aflatoxin B1 detoxification Toxicol Sci 65 35–42 Occurrence Handle10.1093/toxsci/65.1.35 Occurrence Handle1:CAS:528:DC%2BD38XkvFSisA%3D%3D Occurrence Handle11752683

    Article  CAS  PubMed  Google Scholar 

  31. KM Henry CA Townsend (2005) ArticleTitleOrdering the reductive and cytochrome P450 oxidative steps in demethylsterigmatocystin formation yields general insights into the biosynthesis of aflatoxin and related fungal metabolites J Am Chem Soc 127 3724–3733 Occurrence Handle10.1021/ja0455188 Occurrence Handle1:CAS:528:DC%2BD2MXhs1yku7s%3D Occurrence Handle15771506

    Article  CAS  PubMed  Google Scholar 

  32. C Morisseau BL Ward DG Gilchrist BD Hammock (1999) ArticleTitleMultiple epoxide hydrolases in Alternaria alternata f. sp. lycopersici and their relationship to medium composition and host-specific toxin production Appl Environ Microbiol 65 2388–2395 Occurrence Handle1:CAS:528:DyaK1MXjs12gs7c%3D Occurrence Handle10347018

    CAS  PubMed  Google Scholar 

  33. SB Goodwin (2004) ArticleTitleMinimum phylogenetic coverage: an additional criterion to guide the selection of microbial pathogens for initial genomic sequencing efforts Phytopathology 94 800–804 Occurrence Handle18943098

    PubMed  Google Scholar 

  34. UL Rosewich HC Kistler (2000) ArticleTitleRole of horizontal gene transfer in the evolution of fungi Annu Rev Phytopathol 38 325–363 Occurrence Handle10.1146/annurev.phyto.38.1.325 Occurrence Handle1:CAS:528:DC%2BD3cXot1Sju78%3D Occurrence Handle11701846

    Article  CAS  PubMed  Google Scholar 

  35. SE Barnes TP Dola JW Bennett D Bhatnagar (1994) ArticleTitleSynthesis of sterigmatocystin on a chemically defined medium by species of Aspergillus and Chaetomium Mycopathologia 125 173–178 Occurrence Handle10.1007/BF01146523 Occurrence Handle1:CAS:528:DyaK2cXmsFSrtb0%3D Occurrence Handle8047108

    Article  CAS  PubMed  Google Scholar 

  36. YN Han XG Liu U Benny HC Kistler HD VanEtten (2001) ArticleTitleGenes determining pathogenicity to pea are clustered on a supernumerary chromosome in the fungal plant pathogen Nectria haematococca Plant J 25 305–314 Occurrence Handle10.1046/j.1365-313x.2001.00969.x Occurrence Handle1:CAS:528:DC%2BD3MXitlKnu7g%3D Occurrence Handle11208022

    Article  CAS  PubMed  Google Scholar 

  37. PJ Cotty (1994) ArticleTitleInfluence of field application of an atoxigenic strain of Aspergillus flavus on the populations of A. flavus infecting cotton bolls and on the aflatoxin content of cottonseed Phytopathology 84 1270–1277

    Google Scholar 

  38. JW Dorner RJ Cole DT Wicklow (1999) ArticleTitleAflatoxin reduction in corn through field application of competitive fungi J Food Prot 62 650–656 Occurrence Handle1:STN:280:DyaK1MzhtVShug%3D%3D Occurrence Handle10382655

    CAS  PubMed  Google Scholar 

  39. D Bhatnagar KC Ehrlich TE Cleveland (2003) ArticleTitleMolecular genetic analysis and regulation of aflatoxin biosynthesis Appl Microbiol Biotechnol 61 83–93 Occurrence Handle1:CAS:528:DC%2BD3sXit12msbc%3D Occurrence Handle12655449

    CAS  PubMed  Google Scholar 

  40. SP Kale JW Cary D Bhatnagar JW Bennett (1996) ArticleTitleCharacterization of experimentally induced, nonaflatoxigenic variant strains of Aspergillus nidulans Appl Environ Microbiol 62 3399–3404 Occurrence Handle1:CAS:528:DyaK28XlsFeqsbc%3D Occurrence Handle8795232

    CAS  PubMed  Google Scholar 

  41. HH Wilkinson A Ramaswamy SC Sim NP Keller (2004) ArticleTitleIncreased conidiation associated with progression along the sterigmatocystin biosynthetic pathway Mycologia 96 1190–1198 Occurrence Handle1:CAS:528:DC%2BD2MXhsV2ltLc%3D

    CAS  Google Scholar 

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Correspondence to Rosie E. Bradshaw.

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Bradshaw, R.E., Jin, H., Morgan, B.S. et al. A Polyketide Synthase Gene Required for Biosynthesis of the Aflatoxin-like Toxin, Dothistromin. Mycopathologia 161, 283–294 (2006). https://doi.org/10.1007/s11046-006-0240-5

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