Skip to main content
SpringerLink
Log in

TheAspergillus parasiticus polyketide synthase genepksA, a homolog ofAspergillus nidulans wA, is required for aflatoxin B1 biosynthesis

  • Original Paper
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Abstract

Aflatoxins comprise a group of polyketide-derived carcinogenic mycotoxins produced byAspergillus parasiticus andAspergillus flavus. By transformation with a disruption construct, pXX, we disrupted the aflatoxin pathway inA. parasiticus SRRC 2043, resulting in the inability of this strain to produce aflatoxin intermediates as well as a major yellow pigment in the transformants. The disruption was attributed to a single-crossover, homologous integration event between pXX and the recipientA. parasiticus genome at a specific locus, designatedpksA. Sequence analysis suggest thatpksA is a homolog of theAspergillus nidulans wA gene, a polyketide synthase gene involved in conidial wall pigment biosynthesis. The conservedβ-ketoacyl synthase, acyltransferase and acyl carrier-protein domains were present in the deduced amino acid sequence of thepksA product. Noβ-ketoacyl reductase and enoyl reductase domains were found, suggesting thatpksA does not encode catalytic activities for processingβ-carbon similar to those required for long chain fatty acid synthesis. ThepksA gene is located in the aflatoxin pathway gene cluster and is linked to thenor-1 gene, an aflatoxin pathway gene required for converting norsolorinic acid to averantin. These two genes are divergently transcribed from a 1.5 kb intergenic region. We propose thatpksA is a polyketide synthase gene required for the early steps of aflatoxin biosynthesis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Adye, J, Mateles RI (1964) Incorporation of labelled compounds into aflatoxin. Biochim Biophys Acta 86:418–420

    PubMed  Google Scholar 

  • Aguilar F, Harris CC, Sun T, Hollstein M, Cerutti P (1994) Geographic variation of p53 mutational profile in nonmalignant human liver. Science 264:1317–1319

    PubMed  Google Scholar 

  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    PubMed  Google Scholar 

  • Amy CM, Witkowski A, Naggert J, Williams B, Randhawa Z, Smith S (1989) Molecular cloning and sequencing of cDNAs encoding the entire rat fatty acid synthase. Proc Natl Acad Sci USA 86:3114–3118

    PubMed  Google Scholar 

  • Aramayo R, Timberlake WE (1990) Sequence and molecular structure of theAspergillus nidulans yA (laccase I) gene. Nucleic Acids Res 18: 3415

    PubMed  Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1987) Current protocols in molecular biology, vol 1. John Wiley and Sons, New York

    Google Scholar 

  • Beck J, Ripka S, Siegner A, Schiltz E, Schweizer E (1990) The multifunctional 6-methylsalicylic acid synthase gene ofPenicillium patulum. Eur J Biochem 192:487–498

    PubMed  Google Scholar 

  • Bhatnagar D, Cleveland TE, Kingston DGI (1991) Enzymological evidence for separate pathways for aflatoxin B1 and B2 biosynthesis. Biochemistry 30:4343–4350

    PubMed  Google Scholar 

  • Bhatnagar D, Ehrlich KC, Cleveland TE (1992) Oxidation-reduction reactions in biosynthesis of secondary metabolites. In: Bhatnagar D, Lillehoj EB, Arora DK (eds) Handbook of applied mycology, vol 5. Mycotoxins in ecological systems. Marcel Dekker, New York pp 255–286

    Google Scholar 

  • Bhatnagar D, McCormick SP, Lee LS, Hill RA (1987) Identification ofO-methylsterigmatocystin as an aflatoxin B1 and G1 precursor inAspergillus parasiticus. Appl Environ Microbiol 53:1028–1033

    PubMed  Google Scholar 

  • Bressac B, Kew M, Wands J, Ozturk M (1991) Selective G to T mutations of p53 gene in hepatocellular carcinoma from South Africa. Nature 350:429–431

    PubMed  Google Scholar 

  • Brobst SW, Townsend CA (1994) The potential role of fatty acid initiation in the biosynthesis of the fungal aromatic polyketide aflatoxin B1. Can J Chem 72:200–207

    Google Scholar 

  • Chang P-K, Skory CD, Linz JE (1992) Cloning of a gene associated with aflatoxin B1 biosynthesis inAspergillus parasiticus. Curr Genet 21:231–233

    PubMed  Google Scholar 

  • Chang P-K, Bhatnagar D, Cleveland TE (1993a) Disruption of the aflatoxin biosynthetic pathway through genetic complementation inAspergillus parasiticus. FASEB J 7: A1229

    Google Scholar 

  • Chang P-K, Cary JW, Bhatnagar D, Cleveland TE, Bennett JW, Linz JE, Woloshuk CP, Payne GA (1993b) Cloning of theAspergillus parasiticus apa-2 gene associated with the regulation of aflatoxin biosynthesis. Appl Environ Microbiol 59: 3273–3279

    PubMed  Google Scholar 

  • Chirala SS, Kasturi R, Pazirandeh M, Stolow DT, Huang W-Y, Wakil SJ (1989) A novel cDNA extension procedure. J Biol Chem 264:3750–3757

    PubMed  Google Scholar 

  • Cleveland TE, Bhatnagar D, Brown RL (1991) Aflatoxin production via cross-feeding of pathway intermediates during cofermentation of aflatoxin pathway-blockedAspergillus parasiticus mutants. Appl Environ Microbiol 57:2907–2911

    PubMed  Google Scholar 

  • Davis ND, Lyer SK, Diener UL (1987) Improved method of screening for aflatoxin with a coconut agar medium. Appl Environ Microbiol 53:1593–1595

    PubMed  Google Scholar 

  • Donadio S, Staver MJ, McAlpine JB, Swanson S, Katz L (1991) Modular organization of genes required for complex polyketide biosynthesis. Science 252:675–679

    PubMed  Google Scholar 

  • Dutton MF (1988) Enzymes and aflatoxin biosynthesis. Microbiol Rev 53:274–295

    Google Scholar 

  • Eaton DL, Groopman JD (1994) The toxicology of aflatoxins. Academic Press, San Diego

    Google Scholar 

  • Gish W, States DJ (1993) Identification of protein coding regions by database similarity search. Nature Genet 3:266–272

    PubMed  Google Scholar 

  • Heathcote JG, Hibbert JR (1978) Aflatoxins: chemical and biological aspects. Elsevier-Scientific, Amsterdam

    Google Scholar 

  • Hopwood DA, Sherman DH (1990) Molecular genetics of polyketides and its comparison to fatty acid biosynthesis. Annu Rev Genet 24: 37–66

    PubMed  Google Scholar 

  • Horng JS, Chang P-K, Pestka JJ, Linz JE (1990) Development of a homologous transformation system forAspergillus parasiticus with the gene encoding nitrate reductase. Mol Gen Genet 224:294–296

    PubMed  Google Scholar 

  • Hutchinson CR, Decker H, Motamedi H, Shen B, Summers RG (1993) Molecular genetics and biochemistry of a bacterial type II polyketide synthase. In: Baltz RH, Ingolia T, Hageman G (eds) Genetics and molecular biology of industrial microorganisms. American Society for Microbiology, Washington DC, pp 203–216

    Google Scholar 

  • Jelinek CF, Pohland AE, Wood GE (1989) Worldwide occurrence of mycotoxins in foods and feeds—an update. J Assoc Off Anal Chem 72:223–230

    PubMed  Google Scholar 

  • Katz L, Donadio S (1993) Polyketide synthesis: prospects for hybrid antibiotics. Annu Rev Microbiol 47:875–912

    PubMed  Google Scholar 

  • Keller NP, Dischinger HC, Bhatnagar D, Cleveland TE, Ullah AHL (1993) Purification of a 40-kilodalton methyltransferase active in the aflatoxin biosynthetic pathway. Appl Environ Microbiol 59:479–484

    PubMed  Google Scholar 

  • Keller NP, Kantz NJ, Adams TH (1994)Aspergillus nidulans verA is required for production of the mycotoxin sterigmatocystin. Appl Environ Microbiol 60:1444–1450

    PubMed  Google Scholar 

  • Kottig H, Rottner G, Beck K-F, Schweizer M, Schweizer E (1991) The pentafunctionalFASI genes ofSaccharomyces cerevisiae andYarrowia lipolytica are co-linear and considerably longer than previously estimated. Mol Gen Genet 226: 310–314

    PubMed  Google Scholar 

  • Lin BK, Anderson JA (1992) Purification and properties of versicolonal cyclase fromAspergillus parasiticus. Arch Biochem Biophy 293:67–70

    Google Scholar 

  • Mahanti N, Bhatnagar D, Cary J, Linz JE (1995) Structure and function ofuvm8, a gene involved in polyketide backbone synthesis in the aflatoxin pathway inAspergillus parasiticus. Appl Environ Microbiol, in press

  • Mathur M, Kolattukudy PE (1992) Molecular cloning and sequencing of the gene for mycocerosic acid synthase, a novel fatty acid elongation multifunctional enzyme, fromMycobacterium tuberculosis var.bovis Bacillus Calmette-Guerin. J Biol Chem 267:19388–19395

    PubMed  Google Scholar 

  • Mayorga ME, Timberlake WE (1992) The developmentally regulatedAspergillus nidulans wA gene encodes a polypeptide homologous to polyketide and fatty acid synthases. Mol Gen Genet 235:205–212

    PubMed  Google Scholar 

  • Payne GA, Nystorm GJ, Bhatnagar D, Cleveland TE, Woloshuk CP (1993) Cloning of theafl-2 gene involved in aflatoxin biosynthesis fromAspergillus flavus. Appl Environ Microbiol 59:156–162

    PubMed  Google Scholar 

  • Pontecorvo G, Roper JA, Hemmons LM, MacDonald KD, Bufton WJ (1953) The genetics ofAspergillus nidulans. Adv Genet 5:141–238

    PubMed  Google Scholar 

  • Scotti C, Piatti M, Cuzzoni A, Perani P, Tognoni A, Grandi G, Galizzi A, Albertine AM (1993) ABacillus subtilis ORF coding for a polypeptide highly similar to polyketide synthases. Gene 130:65–71

    PubMed  Google Scholar 

  • Skory CD, Chang P-K, Cary J, Linz JE (1992) Isolation and characterization of a gene fromAspergillus parasiticus associated with the conversion of versicolorin A to sterigmatocystin in aflatoxin biosynthesis. Appl Environ Microbiol 58:3527–3537

    PubMed  Google Scholar 

  • Townsend CA, Christensen SB (1983) Stable isotope studies of anthraquinone intermediates in the aflatoxin pathway. Tetrahedron 39:3575–3582

    Google Scholar 

  • Trail F, Chang P-K, Cary J, Linz JE (1994) Structural and functional analysis of thenor-1 gene involved in the biosynthesis of aflatoxins byAspergillus parasiticus. Appl Environ Microbiol 60:4078–4085

    PubMed  Google Scholar 

  • Trail F, Mahanti N, Mehigh R, Liang S-H, Zhou R, Rarich M, Linz JE (1995) A physical and transcriptional map of the aflatoxin gene cluster inAspergillus parasiticus and the functional disruption of a gene involved early in the aflatoxin pathway. Appl Environ Microbiol, in press

  • Vidal-Gros A, Viviani F, Labesse G, Boccara M, Gaudry M (1994) Polyhydroxynaphthalene reductase involved in melanin biosynthesis inMagnaporthe grisea. Eur J Biochem 219:985–992

    PubMed  Google Scholar 

  • Wakil AJ (1989) Fatty acid synthase, a proficient multifunctional enzyme. Biochemistry 28:4523–4530

    PubMed  Google Scholar 

  • Wang IK, Reeves C, Gaucher GM (1991) Isolation and sequencing of a genomic DNA clone containing the 3′ terminus of the 6-methylsalicylic acid polyketide synthetase gene ofPenicillium urticae. Can J Microbiol 37: 86–95

    PubMed  Google Scholar 

  • Yabe K, Ando Y, Hashimoto J, Hamasaki T (1989) Two distinct0-methyltransferases in aflatoxin biosynthesis. Appl Environ Microbiol 55:2172–2177

    PubMed  Google Scholar 

  • Yu J, Cary JW, Bhatnagar D, Cleveland TE, Keller NP, Chu FS (1992) Molecular cloning and characterization of anAspergillus parasiticus gene involved in the conversion of sterigmatocystin to0-methylsterigmatocystin in aflatoxin biosynthesis. Appl Environ Microbiol 59:3564–3571

    Google Scholar 

  • Yu J, Chang P-K, Cary JW, Wright M, Bhatnagar D, Cleveland TE, Payne GA, Linz JE (1995) Comparative mapping of aflatoxin pathway gene clusters inAspergillus parasiticus andAspergillus flavus. Appl Environ Microbiol, 61:2365–2371

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Cell and Molecular Biology, Tulane University, 70118, New Orleans, Louisiana, USA

    Perng-Kuang Chang

  2. Southern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, 1100 Robert E. Lee Boulevard, 70124, New Orleans, Louisiana, USA

    Jeffrey W. Cary, Jiujiang Yu, Deepak Bhatnagar & Thomas E. Cleveland

Authors
  1. Perng-Kuang Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jeffrey W. Cary
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jiujiang Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Deepak Bhatnagar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Thomas E. Cleveland
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chang, PK., Cary, J.W., Yu, J. et al. TheAspergillus parasiticus polyketide synthase genepksA, a homolog ofAspergillus nidulans wA, is required for aflatoxin B1 biosynthesis. Molec. Gen. Genet. 248, 270–277 (1995). https://doi.org/10.1007/BF02191593

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02191593

Key words

Search

Navigation