Characterization of Four Clustered and Coregulated Genes Associated with Fumonisin Biosynthesis in Fusarium verticillioides

Seo, Jeong‐Ah; Proctor, Robert H.; Plattner, Ronald D.

doi:10.1006/fgbi.2001.1299

Cited by 194 publications

(168 citation statements)

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“…Fungal genes encoding enzymes involved in the production of secondary metabolites are usually contained in clusters on chromosomes. The existence of a gene cluster related to mycotoxin synthesis has been demonstrated for aflatoxins, fumonisins, trichothecenes and ergot alkaloids (Yu et al, 2004;Seo et al, 2001;Proctor et al, 2003;Kimura et al, 2007;Panaccione, 2005). By contrast, a gene cluster related to patulin biosynthesis has not been described.…”

Section: Introductionmentioning

confidence: 99%

Molecular cloning and functional characterization of two CYP619 cytochrome P450s involved in biosynthesis of patulin in Aspergillus clavatus

Artigot¹,

Loiseau²,

Laffitte³

et al. 2009

Microbiology

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Patulin is an acetate-derived tetraketide mycotoxin produced by several fungal species, especially Aspergillus, Penicillium and Byssochlamys species. The health risks due to patulin consumption by humans have led many countries to regulate it in human food. Previous studies have shown the involvement of cytochrome P450 monooxygenases in the hydroxylation of two precursors of patulin, m-cresol and m-hydroxybenzylalcohol. In the present study, two cytochrome P450 genes were identified in the genome sequence of Aspergillus clavatus, a patulin-producing species. Both mRNAs were strongly co-expressed during patulin production. CYP619C2, encoded by the first gene, consists of 529 aa, while the second cytochrome, CYP619C3, consists of 524 aa. The coding sequences were used to perform the heterologous expression of functional enzymes in Saccharomyces cerevisiae. The bioconversion assays showed that CYP619C3 catalysed the hydroxylation of m-cresol to yield m-hydroxybenzyl alcohol. CYP619C2 catalysed the hydroxylation of m-hydroxybenzyl alcohol and m-cresol to gentisyl alcohol and 2,5-dihydroxytoluene (toluquinol), respectively. Except for the last compound, all enzyme products are known precursors of patulin. Taken together, these data strongly suggest the involvement of CYP619C2 and CYP619C3 in the biosynthesis of patulin. CYP619C2 and CYP619C3 are located near to two other genes involved in patulin biosynthesis, namely the 6-methylsalicylic acid synthase (6msas) and isoepoxydon dehydrogenase (idh) genes. The current data associated with an analysis of the sequence of A. clavatus suggest the presence of a cluster of 15 genes involved in patulin biosynthesis.

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Section: Introductionmentioning

confidence: 99%

Molecular cloning and functional characterization of two CYP619 cytochrome P450s involved in biosynthesis of patulin in Aspergillus clavatus

Artigot¹,

Loiseau²,

Laffitte³

et al. 2009

Microbiology

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“…Significant progress has been made in elucidating the fumonisin biosynthetic pathway, and regulatory mechanisms associated with the toxin production. The biosynthetic gene cluster, including the polyketide synthase (PKS) gene FUM1 (Proctor et al, 1999(Proctor et al, , 2003Seo et al, 2001), and a number of regulatory genes involved in fumonisin biosynthesis, namely PAC1, FCC1 and ZFR1, have been identified and characterized (Flaherty et al, 2003;Flaherty & Woloshuk, 2004;Shim & Woloshuk, 2001). However, these genes do not show clear epistatic relationships, and therefore it is conceivable that multiple signalling pathways are associated with fumonisin regulation.…”

Section: Introductionmentioning

confidence: 99%

Functional characterization of Fusarium verticillioides CPP1, a gene encoding a putative protein phosphatase 2A catalytic subunit

Choi

Shim

2008

Microbiology

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Fusarium verticillioides produces the mycotoxin fumonisin B 1 (FB 1 ) on maize kernels. In this study, we identified a putative protein phosphatase gene CPP1 in F. verticillioides, and investigated its role in FB 1 regulation. Previous work has shown that CPP1 expression is elevated in an FB 1 -suppressing genetic background. Thus, we hypothesized that CPP1 is negatively associated with FB 1 production. To test this hypothesis, we generated a CPP1 knockout mutant, PP179, and studied the effects of gene deletion on FB 1 biosynthesis and fungal development. PP179 showed elevated expression of FUM genes, and in turn produced higher levels of FB 1 than the wild-type progenitor. Other significant mutant phenotypes included reduced radial growth on agar plates, reduced conidia germination rates, significantly increased macroconidia formation, and hyphal swelling. To verify that these phenotypes were directly due to CPP1 deletion, we complemented PP179 with the wild-type CPP1 gene. The complemented strain PPC4 showed FUM1 expression and FB 1 production similar to that of the wild-type, providing evidence that CPP1 is negatively associated with FB 1 biosynthesis. Other PP179 phenotypes, such as macroconidiation and hyphal swelling, were also restored to that of wild-type progenitor. Furthermore, we complemented F. verticillioides PP179 strain with Neurospora crassa wild-type ppe-1 gene, demonstrating that Cpp1 and PPE-1 proteins are functionally conserved. Pleiotropic effects of CPP1 deletion led us to hypothesize that CPP1 is associated with multiple downstream signalling pathways in F. verticillioides. Identification and functional characterization of downstream Cpp1-interacting proteins are necessary to better understand the complex regulatory mechanisms associated with Cpp1.

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“…Clustering is not observed for most biosynthetic genes in the SM of higher eukaryotes, with a few exceptions in plants (7,21). In organisms like filamentous fungi, such clustering occurs and is maintained probably because it facilitates horizontal transfer and coordinated transcriptional regulation of the genes therein (29).The FB biosynthetic gene cluster consists of 17 transcriptionally coregulated genes designated FUM1 through FUM3 and FUM6 through FUM21 (with FUM2 and FUM3 later found to be the same as FUM12 and FUM9, respectively) (1,5,18,26). While the functions of some of these genes are only hypothesized, others have been characterized experimentally.…”

mentioning

confidence: 99%

“…The FB biosynthetic gene cluster consists of 17 transcriptionally coregulated genes designated FUM1 through FUM3 and FUM6 through FUM21 (with FUM2 and FUM3 later found to be the same as FUM12 and FUM9, respectively) (1,5,18,26). While the functions of some of these genes are only hypothesized, others have been characterized experimentally.…”

mentioning

confidence: 99%

“…In particular, FUM1 plays a key role in biosynthesis because its product is a polyketide synthase involved at an early step in the assembly of the FB backbone (19). FUM8 encodes an aminoacyl transferase that catalyzes the formation of the full-length C 20 molecule by adding a carbanion derived from the ␣-carbon of alanine to the polyketide backbone generated by Fum1; the disruption of these two genes blocks FB production (1,26).…”

mentioning

confidence: 99%

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Transcription of Genes in the Biosynthetic Pathway for Fumonisin Mycotoxins Is Epigenetically and Differentially Regulated in the Fungal Maize Pathogen Fusarium verticillioides

et al. 2012

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When the fungal pathogen Gibberella moniliformis (anamorph, Fusarium verticillioides) colonizes maize and maize-based products, it produces class B fumonisin (FB) mycotoxins, which are a significant threat to human and animal health. FB biosynthetic enzymes and accessory proteins are encoded by a set of clustered and cotranscribed genes collectively named FUM, whose molecular regulation is beginning to be unraveled by researchers. FB accumulation correlates with the amount of transcripts from the key FUM genes, FUM1, FUM21, and FUM8. In fungi in general, gene expression is often partially controlled at the chromatin level in secondary metabolism; when this is the case, the deacetylation and acetylation (and other posttranslational modifications) of histones are usually crucial in the regulation of transcription. To assess whether epigenetic factors regulate the FB pathway, we monitored FB production and FUM1, FUM21, and FUM8 expression in the presence of a histone deacetylase inhibitor and verified by chromatin immunoprecipitation the relative degree of histone acetylation in the promoter regions of FUM1, FUM21, and FUM8 under FB-inducing and noninducing conditions. Moreover, we generated transgenic F. verticillioides strains expressing GFP under the control of the FUM1 promoter to determine whether its strength under FB-inducing and noninducing conditions was influenced by its location in the genome. Our results indicate a clear and differential role for chromatin remodeling in the regulation of FUM genes. This epigenetic regulation can be attained through the modulation of histone acetylation at the level of the promoter regions of the key biosynthetic genes FUM1 and FUM21, but less so for FUM8.F umonisins are a family of mycotoxins produced by the secondary metabolism (SM) of Fusarium verticillioides (teleomorph, Gibberella moniliformis) and Fusarium proliferatum that contaminate maize and maize-based products. Within the B series of these toxins (FB), FB1, FB2, and FB3 are the ones most frequently found under field conditions and have been linked to various animal and human mycotoxicoses (23). FB are polyketides consisting of a linear 19-or 20-carbon backbone with hydroxyl, methyl, and tricarballylic acid moieties at various positions along the base chain (19). In filamentous Ascomycetes, genes involved in the biosynthesis of toxins (such as aflatoxins and trichothecenes) and of other secondary metabolites are frequently organized into clusters (6). Clustering is not observed for most biosynthetic genes in the SM of higher eukaryotes, with a few exceptions in plants (7,21). In organisms like filamentous fungi, such clustering occurs and is maintained probably because it facilitates horizontal transfer and coordinated transcriptional regulation of the genes therein (29).The FB biosynthetic gene cluster consists of 17 transcriptionally coregulated genes designated FUM1 through FUM3 and FUM6 through FUM21 (with FUM2 and FUM3 later found to be the same as FUM12 and FUM9, respectively) (1,5,18,26). While the func...

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Characterization of Four Clustered and Coregulated Genes Associated with Fumonisin Biosynthesis in Fusarium verticillioides

Cited by 194 publications

References 44 publications

Molecular cloning and functional characterization of two CYP619 cytochrome P450s involved in biosynthesis of patulin in Aspergillus clavatus

Molecular cloning and functional characterization of two CYP619 cytochrome P450s involved in biosynthesis of patulin in Aspergillus clavatus

Functional characterization of Fusarium verticillioides CPP1, a gene encoding a putative protein phosphatase 2A catalytic subunit

Transcription of Genes in the Biosynthetic Pathway for Fumonisin Mycotoxins Is Epigenetically and Differentially Regulated in the Fungal Maize Pathogen Fusarium verticillioides

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