Global phenotypic analysis and transcriptional profiling defines the weak acid stress response regulon in Saccharomyces cerevisiae

26 2006

Global phenotypic analysis and transcriptional profiling defines the weak acid stress response regulon in Saccharomyces cerevisiae

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Christoph Schüller1, Yasmine M. Mamnun1, Mehdi Mollapour2, Gerd Krapf1, Michael Schuster1, Bettina E. Bauer1, Peter W. Piper2, and Karl Kuchler1*

1 Institute of Medical Biochemistry, Department of Molecular Genetics, University and BioCenter of Vienna, A-1030 Vienna, Austria
2 Department of Biochemistry and Molecular Biology, University College London, London WC1E 6BT, U.K.

* Corresponding author. E-mail address: .

Weak organic acids such as sorbate are potent fungistatic agents used in food preservation, but their intracellular targets are poorly understood. We thus searched for potential target genes and signaling components in the yeast genome using contemporary genome-wide functional assays, as well as DNA microarray profiling.

Phenotypic screening of the EUROSCARF collection revealed the existence of numerous sorbate-sensitive strains. Sorbate hypersensitivity was detected in mutants of the shikimate biosynthesis pathway, strains lacking the PDR12 efflux pump or WAR1, a transcription factor mediating stress induction of PDR12. Using DNA microarrays, we also analyzed the genome-wide response to acute sorbate stress, allowing for the identification of more than 100 genes rapidly induced by weak acid stress.

Morever, a novel War1p and Msn2p/4p-independent regulon which includes HSP30 was identified. Although induction of the majority of sorbate-induced genes required Msn2p/4p, weak acid tolerance was unaffected by a lack of Msn2p/4p.

Ectopic expression of PDR12 from the GAL1-10 promoter fully restored sorbate resistance in a strain lacking War1p, demonstrating that PDR12 is the major target of War1p under sorbic acid stress. Interestingly, comparison of microarray data with results from the phenotypic screening revealed that PDR12 remained as the only gene, which is both stress-inducible and required for weak acid resistance.

Our results suggest that combining functional assays with transcriptome profiling allows for the identification of key components in large datasets such as those generated by global microarray analysis.

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