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Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans
Genetics and Developmental Biology Program, Division of Plant and Soil Sciences, West Virginia University, Morgantown, WV 26506, USA
* Author to whom correspondence should be addressed.
Received: 23 January 2013; in revised form: 8 February 2013 / Accepted: 17 February 2013 / Published: 22 February 2013
Abstract: 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.
Keywords: ergot alkaloids; mycotoxins; Aspergillus fumigatus; gene cluster; chanoclavine-I
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Ryan, K.L.; Moore, C.T.; Panaccione, D.G. Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans. Toxins 2013, 5, 445-455.
Ryan KL, Moore CT, Panaccione DG. Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans. Toxins. 2013; 5(2):445-455.
Ryan, Katy L.; Moore, Christopher T.; Panaccione, Daniel G. 2013. "Partial Reconstruction of the Ergot Alkaloid Pathway by Heterologous Gene Expression in Aspergillus nidulans." Toxins 5, no. 2: 445-455.