Aflatoxin biosynthesis and regulation
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1 Aflatoxin biosynthesis and regulation Kenneth Ehrlich Southern Regional Research Center New rleans
2 Introduction AF/ST biosynthesis involves expression of at least 25 genes Genes are organized in a cluster or clusters Genes are coordinately regulated The pathway-specific transcription factor, AflR, and globally acting transcription factors regulate biosynthesis Regulation of biosynthesis is different in different AF-producing species
3 Comparison of AF/ST clusters omtb norb cypa aflt pksa hypb1 nor-1 hexa hexb aflr aflj adha esta nora ver-1 vera avna verb hypb2 - avfa omtb omta orda vbs cypx moxy ordb hypa 1 nada hypa AF cluster ST cluster A. ochraceoros. AF clusters DNA identity (%) stcs stca B CD E aflraflj F G H I J K L M N P Q R S T U V W A B CD E aflraflj F G H I J K L M N P Q R T U V W D. septosporum dothistromin clusters dotd C B A pksa cypx TE transp ver-1 avfa moxy epoa hexa vbs
4 Polyketide synthesis HexA HexB ACP KR KS PPT AT ER ER FMH NADH DH MAT PksA SAT KS MAT PT ACP TE/CLC ACP KS MAT PPT SAT PT TE/CLC ER DH KR AT acyl carrier protein keto synthase malonylcoa ACPtransacylase phosphopantetheinyltransferase starter unit ACP transacylase product template thioesterase/claisen-like cyclase enoylreductase dehydratase ketoreductase acetyltransacylase
5 NorS complex S 2 x MalonylCoA S HexA HexB PksA HexA HexB PksA H H SH S 7 x MalonylCoA HexA HexB PksA H Noranthrone H
6 Watanabe&Ebizuka 2004 Control of PKS size by CLC domain
7 verview of AflR and aflr intergenic region Promoter with transcription factor binding sites aflj BrlA AflR AflR AbaA AbaA BrlA tsp aflr gene G1-3 G4,5 AreA-binding sites aflj\aflr Intergenic region Zn cluster His-repeat Pro-rich PEST domains Acidic domain Ser-rich His-Arg NLE Linker AflR protein
8 Variability in putative transcription factor binding sites in three AF cluster genes AP AN A.nid 50bp 100bp aflr-aflj intergenic region omta-orda intergenic region TATA CAAT AflR AreA PacC BrlA AbaA CreA 100bp omtb promoter; omta-omtb intergenic region AP AN / /
9 AflR coding domain variability Strain His-rich region (No. of His) PEST region A. flavus (Grp I) 6 No A. parasit. 6 No A. flavus (Grp II) 2 No A. pseudotamar. 6 No A. nomius 4 Yes A. bombycis 4 Yes A. nidulans A. ochraceo. 0 Yes 0 Yes
10 ther levels of regulation LaeA protein methyltransferase: affects only 2º metabolite clusters; may specifically affect pre mrna stability, translation PKA VeA mediated by G-protein signaling factors: FlbA and FluG possible transcription factor: mediates formation of both sclerotia and AF; target is unknown.
11 Epigenetic regulation Chromosomal position of gene cluster DNA methylation of GC-rich regions in subtelomeric region Histone modifications: acetylation, methylation; phosphorylation
12 Conclusions Polyketide formation utilizes a Fas-derived starter unit and an iterative PKS Regulation of AF biosynthesis involves AflR and global transcription factors Some biosynthesis steps involve several oxidative enzymes, but no stable intermediates are detectable
13
14 Conversion of Avf to HVN H H H CypX H AvfA CH averufin H H H H H hydroxyversicolorone
15 Conversion of VerA to ST H H H H H A B C D E VerA H AflX VA I H H H H H II Ver-1 III AflY H H H H C 2 H H IV -C 2 -H 2 DMST mtb ST
16 AF formation VerA mtb, mta Me rda Me NIH shift Me H rda Me Me Sterigmatocystin Me H Me NorA Aflatoxin B1 HypB2 Me H 2 C CypA Me NadA NorB Aflatoxin G1 H 2 C
Clustered Pathway Genes in Aflatoxin Biosynthesis
REFERENCES CONTENT ALERTS Clustered Pathway Genes in Aflatoxin Biosynthesis Jiujiang Yu, Perng-Kuang Chang, Kenneth C. Ehrlich, Jeffrey W. Cary, Deepak Bhatnagar, Thomas E. Cleveland, Gary A. Payne, John
Aflatoxin biosynthesis
Review Rev Iberoam Micol 2002; 19: 191-200 191 Aflatoxin biosynthesis Jiujiang Yu, Deepak Bhatnagar & Kenneth C. Ehrlich United States Department of Agriculture, Agricultural Research Service, Southern
Conservation, University of Bonn, Bonn, Germany b International Institute of Tropical Agriculture (IITA), PMB 5320, Ibadan, Nigeria
This article was downloaded by: [University of Arizona] On: 11 August 2014, At: 15:56 Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office:
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