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Characterization of the Ustilago maydis sid2 Gene, Encoding a Multidomain Peptide Synthetase in the Ferrichrome Biosynthetic Gene Cluster

Yuan, Walter M. and Gentil, Guillaume D. and Budde, Allen D. and Leong, Sally A. (2001) Characterization of the Ustilago maydis sid2 Gene, Encoding a Multidomain Peptide Synthetase in the Ferrichrome Biosynthetic Gene Cluster. Journal of Bacteriology, 183 (13). pp. 4040-4051. ISSN 0021-9193. https://resolver.caltech.edu/CaltechAUTHORS:YUAjbact01

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Abstract

Ustilago maydis, the causal agent of corn smut disease, acquires and transports ferric ion by producing the extracellular, cyclic peptide, hydroxamate siderophores ferrichrome and ferrichrome A. Ferrichrome biosynthesis likely proceeds by hydroxylation and acetylation of L-ornithine, and later steps likely involve covalently bound thioester intermediates on a multimodular, nonribosomal peptide synthetase. sid1 encodes L-ornithine N5-oxygenase, which catalyzes hydroxylation of L-ornithine, the first committed step of ferrichrome and ferrichrome A biosynthesis in U. maydis. In this report we characterize sid2, another biosynthetic gene in the pathway, by gene complementation, gene replacement, DNA sequence, and Northern hybridization analysis. Nucleotide sequencing has revealed that sid2 is located 3.7 kb upstream of sid1 and encodes an intronless polypeptide of 3,947 amino acids with three iterated modules of an approximate length of 1,000 amino acids each. Multiple motifs characteristic of the nonribosomal peptide synthetase protein family were identified in each module. A corresponding iron-regulated sid2 transcript of 11 kb was detected by Northern hybridization analysis. By contrast, constitutive accumulation of this large transcript was observed in a mutant carrying a disruption of urbs1, a zinc finger, GATA family transcription factor previously shown to regulate siderophore biosynthesis in Ustilago. Multiple GATA motifs are present in the intergenic region between sid1 and sid2, suggesting bidirectional transcription regulation by urbs1 of this pathway. Indeed, mutation of two of these motifs, known to be important to regulation of sid1, altered the differential regulation of sid2 by iron.


Item Type:Article
Additional Information:Copyright © 2001, American Society for Microbiology. Received 20 December 2000/Accepted 10 April 2001 We are very grateful to ZhiQiang An and Hans von Doehren for their helpful discussions and critical review of the manuscript and Gunther Winkelmann for providing the figure of the structure of the ferrichromes. This work was supported by the USDA-ARS and NIH grant GM33716 to S.A.L. M. W. Yuan and Guillaume D. Gentil contributed equally to the work described in this paper.
Issue or Number:13
Record Number:CaltechAUTHORS:YUAjbact01
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:YUAjbact01
Alternative URL:http://dx.doi.org/10.1128/JB.183.13.4040-4051.2001
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:2210
Collection:CaltechAUTHORS
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Deposited On:15 Mar 2006
Last Modified:02 Oct 2019 22:51

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