Subtle hydrophobic interactions between the seventh residue of the zinc finger loop and the first base of an HGATAR sequence determine promoter-specific recognition by the Aspergillus nidulans GATA factor AreA

Abstract

A change of a universally conserved leucine to valine in the DNA‐binding domain of the GATA factor AreA results in inability to activate some AreA‐dependent promoters, including that of the uapA gene encoding a specific urate–xanthine permease. Some other AreA‐ dependent promoters become able to function more efficiently than in the wild‐type context. A methionine in the same position results in a less extreme, but opposite effect. Suppressors of the AreA(Val) mutation mapping in the uapA promoter show that the nature of the base in the first position of an HGATAR (where H stands for A, T or C) sequence determines the relative affinity of the promoter for the wild‐type and mutant forms of AreA. In vitro binding studies of wild‐type and mutant AreA proteins are completely consistent with the phenotypes in vivo. Molecular models of the wild‐type and mutant AreA–DNA complexes derived from the atomic coordinates of the GATA‐1–AGATAA complex account both for the phenotypes observed in vivo and the binding differences observed in vitro. Our work extends the consensus of physiologically relevant binding sites from WGATAR to HGATAR, and provides a rationale for the almost universal evolutionary conservation of leucine at the seventh position of the Zn finger of GATA factors. This work shows inter alia that the sequence CGATAGagAGATAA, comprising two almost adjacent AreA‐binding sites, is sufficient to ensure activation of transcription of the uapA gene.

Nitrogen Metabolite Repression of Metabolism and Virulence in the Human Fungal Pathogen Cryptococcus neoformans

“…Bwc2, Cir1, Gat201, and Gat1/Are1 together with the three other uncharacterized GATA factors (CNAG03401.2, CNAG04263.2, CNAG06762.2) we have identified are predicted to bind to 59-GATA-39 sites in the genome (Ko and Engel 1993;Merika and Orkin 1993;Ravagnani et al 1997). Whether these GATA factors antagonize and compete for the same 59-GATA-39 binding sites in the promoter regions of genes, act in synergy to activate transcription of genes, or act interchangably remains to be determined.…”

“…These proteins are distinguished by their highly conserved DNA-binding domain consisting of a Cys 2 /Cys 2 -type zinc finger motif followed by an adjacent basic region and are named after the DNA sequence 59-GATA-39 that they recognize (Ko and Engel 1993;Merika and Orkin 1993;Ravagnani et al 1997). GATA transcription factors are widely found in eukaryotes, including metazoans and plants, and have diverse biological functions.…”

Sulphur and nitrogen regulation of the protease-encoding ACP1 gene in the fungus Botrytis cinerea: correlation with a phospholipase D activity

“…The consensus 'agataa' sequence for AREA and 'atgtcggcat' sequence for CYS3 were changed to 'agggaa' and 'atgtggcat', respectively. These mutations had been shown to disrupt the nitrogen and sulphur regulation of other genes (Li & Marzluf, 1996;Ravagnani et al, 1997).…”

Global nutrient profiling by Phenotype MicroArrays: a tool complementing genomic and proteomic studies in conidial fungi

“…These factors have been intensively studied in A. nidulans, N. crassa, H. jecorina, and Acremonium chrysogenum (Bailey and Arst, 1975;Kudla et al, 1990;Dowzer and Kelly, 1991;Caddick et al, 1994;Strauss et al, 1995;Ilmén et al, 1996;Platt et al, 1996;Ravagnani et al, 1997;Marzluf, 1997;Wilson and Arst, 1998;Jekosch and Kück, 2000;Felenbok et al, 2001). In this type of experiment, growth of a parent strain and a respective mutant would be compared on every medium composition, which would reveal those particular nutrients whose utilization depends on the mutated wide-domain control gene.…”

“…Mutations in areA exhibit an extraordinarily diverse range of phenotypes when monitored for their effects on utilization of nitrogenous compounds. One of these alleles, areA-102, displays altered specificity of target promoter activation, which ultimately leads to loss-of-function, gain-of-function, or a wild-type phenotype based on its interaction with the target structural gene promoter (Kudla et al, 1990;Ravagnani et al, 1997). In comparison to wild-type strains on agar plates, strains carrying areA-102 show increased growth when urea, L-histidine, L-citrulline, L-aspartate, or L-glutamate is used as the sole nitrogen source and decreased growth when xanthine or uric acid is used as the sole nitrogen source (Arst and Cove, 1973; Arst and Scazzocchio, 1975;Hynes, 1975;Kudla et al, 1990).…”