Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans

Ergot alkaloids are pharmaceutically and agriculturally important secondary metabolites produced by several species of fungi. Ergot alkaloid pathways vary among different fungal lineages, but the pathway intermediate chanoclavine-I is evolutionarily conserved among ergot alkaloid producers. At least...

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Main Authors: Daniel G. Panaccione, Christopher T. Moore, Katy L. Ryan
Format: Article
Language:English
Published: MDPI AG 2013-02-01
Series:Toxins
Subjects:
Online Access:http://www.mdpi.com/2072-6651/5/2/445
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author Daniel G. Panaccione
Christopher T. Moore
Katy L. Ryan
author_facet Daniel G. Panaccione
Christopher T. Moore
Katy L. Ryan
author_sort Daniel G. Panaccione
collection DOAJ
description Ergot alkaloids are pharmaceutically and agriculturally important secondary metabolites produced by several species of fungi. Ergot alkaloid pathways vary among different fungal lineages, but the pathway intermediate chanoclavine-I is evolutionarily conserved among ergot alkaloid producers. At least four genes, dmaW, easF, easE, and easC, are necessary for pathway steps prior to chanoclavine-I; however, the sufficiency of these genes for chanoclavine-I synthesis has not been established. A fragment of genomic DNA containing dmaW, easF, easE, and easC was amplified from the human-pathogenic, ergot alkaloid-producing fungus Aspergillus fumigatus and transformed into Aspergillus nidulans, a model fungus that does not contain any of the ergot alkaloid synthesis genes. HPLC and LC-MS analyses demonstrated that transformed A. nidulans strains produced chanoclavine-I and an earlier pathway intermediate. Aspergillus nidulans transformants containing dmaW, easF, and either easE or easC did not produce chanoclavine-I but did produce an early pathway intermediate and, in the case of the easC transformant, an additional ergot alkaloid-like compound. We conclude that dmaW, easF, easE, and easC are sufficient for the synthesis of chanoclavine-I in A. nidulans and expressing ergot alkaloid pathway genes in A. nidulans provides a novel approach to understanding the early steps in ergot alkaloid synthesis.
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spelling doaj.art-b157bdea4f8d46b39842455e12429f9e2022-12-22T04:21:13ZengMDPI AGToxins2072-66512013-02-015244545510.3390/toxins5020445Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulansDaniel G. PanaccioneChristopher T. MooreKaty L. RyanErgot alkaloids are pharmaceutically and agriculturally important secondary metabolites produced by several species of fungi. Ergot alkaloid pathways vary among different fungal lineages, but the pathway intermediate chanoclavine-I is evolutionarily conserved among ergot alkaloid producers. At least four genes, dmaW, easF, easE, and easC, are necessary for pathway steps prior to chanoclavine-I; however, the sufficiency of these genes for chanoclavine-I synthesis has not been established. A fragment of genomic DNA containing dmaW, easF, easE, and easC was amplified from the human-pathogenic, ergot alkaloid-producing fungus Aspergillus fumigatus and transformed into Aspergillus nidulans, a model fungus that does not contain any of the ergot alkaloid synthesis genes. HPLC and LC-MS analyses demonstrated that transformed A. nidulans strains produced chanoclavine-I and an earlier pathway intermediate. Aspergillus nidulans transformants containing dmaW, easF, and either easE or easC did not produce chanoclavine-I but did produce an early pathway intermediate and, in the case of the easC transformant, an additional ergot alkaloid-like compound. We conclude that dmaW, easF, easE, and easC are sufficient for the synthesis of chanoclavine-I in A. nidulans and expressing ergot alkaloid pathway genes in A. nidulans provides a novel approach to understanding the early steps in ergot alkaloid synthesis.http://www.mdpi.com/2072-6651/5/2/445ergot alkaloidsmycotoxinsAspergillus fumigatusgene clusterchanoclavine-I
spellingShingle Daniel G. Panaccione
Christopher T. Moore
Katy L. Ryan
Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans
Toxins
ergot alkaloids
mycotoxins
Aspergillus fumigatus
gene cluster
chanoclavine-I
title Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans
title_full Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans
title_fullStr Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans
title_full_unstemmed Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans
title_short Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans
title_sort partial reconstruction of the ergot alkaloid pathway by heterologous gene expression in aspergillus nidulans
topic ergot alkaloids
mycotoxins
Aspergillus fumigatus
gene cluster
chanoclavine-I
url http://www.mdpi.com/2072-6651/5/2/445
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AT christophertmoore partialreconstructionoftheergotalkaloidpathwaybyheterologousgeneexpressioninaspergillusnidulans
AT katylryan partialreconstructionoftheergotalkaloidpathwaybyheterologousgeneexpressioninaspergillusnidulans