Sequences and proteins required for iron-regulated expression ofsid1ofUstilago maydis

Abstract

The molecular biology of the high affinity, siderophore-mediated iron uptake system of the basidiomycete fungus Ustilago maydis is under investigation. Ustilago maydis produces two cyclic peptide siderophores, ferrichrome and ferrichrome A. Biosynthesis of both siderophores is initiated by ornithine-N5-oxygenase, the product of sid1. sid1 mRNA accumulates only during growth under iron starvation conditions in wild-type cells or constitutively in urbs1 mutants, urbs1 encodes a 100-kDa protein with putative Zn finger domains that share sequence identity with those of the GATA family of transcription factors. The promoter region of sid1 was defined by deletion analysis of a 3.0-kb region 5′ to the translational start of sid1 using the Escherichia coli GUS gene as a reporter. Three regions were defined by this analysis to be critical to expression of sid1. These include (i) a 306-bp region containing two GATA sequences and mapping 2.4 kb from the start of translation; (ii) a 439-bp region immediately 5′ to the start of transcription; and (iii) a region encompassing the first intron of sid1. Deletion of the GATA sequences resulted in deregulated expression of sid1, while elimination of the latter two sequences ablated expression of the gene under all circumstances. Current efforts are focused on determining whether Urbsl interacts directly with the sid1 promoter via the GATA sequences and whether this interaction is dependent upon iron. Key words: GATA, transcription factor, siderophore, ferrichrome, iron, Urbs1.