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Reference: Gregori C, et al. (2008) Weak organic acids trigger conformational changes of the yeast transcription factor war1 in vivo to elicit stress adaptation. J Biol Chem 283(37):25752-64

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Abstract

The Saccharomyces cerevisiae zinc cluster regulator War1 mediates an essential transcriptional and adaptive response to weak organic acid stress. Here, we investigate the mechanism of War1 activation upon weak acid stress. We identified several gain-of-function WAR1 alleles mapping to the central War1 region. These mutations constitutively increase levels of the plasma membrane ABC transporter Pdr12, the main War1 target mediating stress adaptation. Functional analysis of War1 reveals that the central region and its C-terminal activation domain are required for function. Notably, the native DNA-binding and dimerization domains appear dispensable for War1 activity, since they can be replaced by a LexA DNA-binding domain. Chromatin immunoprecipitation (ChIP) demonstrates elevated promoter affinity of activated War1, since its PDR12 promoter association increases upon stress. Hyperactive WAR1 alleles have constitutively high PDR12 promoter association. Furthermore, fluorescence resonance energy transfer (FRET) of functional CFP-War1-YFP proteins also demonstrates conformational changes of stress-activated War1 in vivo. Our results suggest a mechanism whereby War1 activation is accompanied by conformational changes enhancing promoter association, thus initiating the adaptation process.

Reference Type
Journal Article
Authors
Gregori C, Schuller C, Frohner IE, Ammerer G, Kuchler K
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