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Identification of an Hsp90 mutation that selectively disrupts cAMP/PKA signaling in Saccharomyces cerevisiae

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Abstract

The molecular chaperone Hsp90 cooperates with multiple cochaperone proteins as it promotes the folding and activation of diverse client proteins. Some cochaperones regulate the ATPase activity of Hsp90, while others appear to promote Hsp90 interaction with specific types of client proteins. Through its interaction with the adenylate cyclase Cyr1, the Sgt1 cochaperone modulates the activity of the cAMP pathway in Saccharomyces cerevisiae. A specific mutation in yeast Hsp90, hsc82-W296A, or a mutation in Sgt1, sgt1-K360E, resulted in altered transcription patterns genetically linked to the cAMP pathway. Hsp90 interacted with Cyr1 in vivo and the hsc82-W296A mutation resulted in reduced accumulation of Cyr1. Hsp90-Sgt1 interaction was altered by either the hsc82-W296A or sgt1-K360E mutation, suggesting defective Hsp90-Sgt1 cooperation leads to reduced Cyr1 activity. Microarray analysis of hsc82-W296A cells indicated that over 80 % of all transcriptional changes in this strain may be attributed to altered cAMP signaling. This suggests that a majority of the cellular defects observed in hsc82-W296A cells are due to altered interaction with one specific essential cochaperone, Sgt1 and one essential client, Cyr1. Together our results indicate that specific interaction of Hsp90 and Sgt1 with Cyr1 plays a key role in regulating gene expression, including genes involved in polarized morphogenesis.

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Acknowledgments

We thank Karen Miller and Gustavo Arrizabalaga for assistance with RNA isolation and QTPCR. We also thank Colby Austin, Derek Pierson and Adam Theun for assistance with plasmid and strain construction, and Abbey Zuehlke for technical assistance and careful reading of the manuscript. We would like to thank Matthew Settles and Derek Pouchnik for their assistance with the microarray and analysis of the microarray data. The antibody against Tim44 was a generous gift from Dr. Elizabeth Craig (University of Wisconsin-Madison). This study was supported by Grant MCB-0744522 from the National Science Foundation.

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  1. Department of Biological Sciences, Center for Reproductive Biology, University of Idaho, Life Sciences South Room 252, P.O. Box 443051, Moscow, ID, 83844-3051, USA

    Gary A. Flom, Ewa Langner & Jill L. Johnson

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  1. Gary A. Flom
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  2. Ewa Langner
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Correspondence to Jill L. Johnson.

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Flom, G.A., Langner, E. & Johnson, J.L. Identification of an Hsp90 mutation that selectively disrupts cAMP/PKA signaling in Saccharomyces cerevisiae . Curr Genet 58, 149–163 (2012). https://doi.org/10.1007/s00294-012-0373-7

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