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Activation of the Ustilagic Acid Biosynthesis Gene Cluster in Ustilago maydis by the C₂H₂ Zinc Finger Transcription Factor Rua1
- Teichmann, Beate, Liu, Lidan, Schink, Kay Oliver, Bölker, Michael
- Applied and environmental microbiology 2010 v.76 no.8 pp. 2633-2640
- DNA, Ustilago zeae, acids, binding sites, biological control, biosurfactants, biosynthesis, cellobiose, consensus sequence, conserved sequences, multigene family, nitrogen, nuclear proteins, plant pathogenic fungi, starvation, structural genes, toxicity, transcription factors, yeasts, zinc finger motif
- The phytopathogenic basidiomycetous fungus Ustilago maydis secretes, under conditions of nitrogen starvation, large amounts of the biosurfactant ustilagic acid (UA). This secreted cellobiose glycolipid is toxic for many microorganisms and confers biocontrol activity to U. maydis. Recently, a large gene cluster that is responsible for UA biosynthesis was identified. Here, we show that expression of all cluster genes depends on Rua1, a nuclear protein of the C₂H₂ zinc finger family, whose gene is located within the gene cluster. While deletion of rua1 results in complete loss of UA production, overexpression of rua1 promotes increased UA synthesis even in the presence of a good nitrogen source. Bioinformatic analysis allowed us to identify a conserved sequence element that is present in the promoters of all structural genes involved in UA biosynthesis. Deletion analysis of several promoters within the cluster revealed that this DNA element serves as an upstream activating sequence (UAS) and mediates Rua1-dependent expression. We used the yeast one-hybrid system to demonstrate specific recognition of this DNA element by Rua1. Introduction of nucleotide exchanges into the consensus sequence interfered with Rua1-dependent activation, suggesting that this sequence element acts as a direct binding site for Rua1.