Abstract
In Saccharomyces cerevisiae cells, high external osmolarity leads to the activation of a p38-related mitogen-activated protein (MAP) kinase though Pbs2. Pbs2 tagged with green fluorescent protein (Pbs2–GFP) is evenly distributed in the cytoplasm but excluded from the nucleus before and after exposure to stress. Here we show that a catalytically inactive form of Pbs2 attains a highly polarised localization during osmostress. This phenomenon depends of the osmosensor Sho1 and on a functional Cdc42 GTPase. Cdc42, but not the actin cytoskeleton, influences Sho1-dependent activation of the MAP kinase. Sho1 itself accumulates at sites of polar growth, but independently of stress conditions and Cdc42. These observations allow us to define the sequence of events that occurs during propogation of osmostress signals.
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Acknowledgements
We thank D. Drubin, K. Nasmyth, M. Ramezani Rad, S. O'Rourke and H. Saito for strains and plasmids, H. Ruis and all the members of the Ruis and Ammerer laboratories for discussions and material support, and L. Huber and P. Kovarik for comments on the manuscript. We are greatful to S. Reipert for help with the laser-scanning microscope. Latrunculin A was kindly supplied by M. Sanders. This work was partly supported by TMR Network grant ERBFMRX-CT96-0041 (to G.A.) and by student program grant W001 from the Austrian Fonds zur Förderung Wissenschaftlicher Forschung.
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Reiser, V., Salah, S. & Ammerer, G. Polarized localization of yeast Pbs2 depends on osmostress, the membrane protein Sho1 and Cdc42. Nat Cell Biol 2, 620–627 (2000). https://doi.org/10.1038/35023568
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DOI: https://doi.org/10.1038/35023568
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