Abstract
Numerous putative heat shock protein 90 (Hsp90)-interacting proteins, which could represent novel folding clients or co-chaperones, have been identified in recent years. Two separate high-throughput screens in yeast uncovered genetic effects between Hsp90 and components of the ER membrane complex (EMC), which is required for tolerance to unfolded protein response stress in yeast. Herein, we provide the first experimental evidence supporting that there is a genuine interaction of Hsp90 with the EMC. We demonstrate genetic interactions between EMC2 and the known Hsp90 co-chaperone encoded by STI1, as well as Hsp90 point mutant allele-specific differences in inherent growth and Hsp90 inhibitor tolerance in the absence and presence of EMC2. In co-precipitation experiments, Hsp90 interacts with Emc2p, whether or not Emc2p contains amino acid sequences designated as a tetratricopeptide repeat motif. Yeast with multiple EMC gene deletions exhibit increased sensitivity to Hsp90 inhibitor as well as defective folding of the well-established Hsp90 folding client, the glucocorticoid receptor. Altogether, our evidence of physical, genetic, and functional interaction of Hsp90 with the EMC, as well as bioinformatic analysis of shared interactors, supports that there is a legitimate interaction between them in vivo.
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Acknowledgements
We are grateful to Jill Johnson for Hsp90 co-chaperone expression plasmids, as well as v-src, Ste11, and glucocorticoid receptor folding assay plasmids, and to William Prinz for supplying the multiple EMC deletion strains. We additionally thank David Norman and Gerry Bolgos for their general laboratory assistance. A.J.M. thanks Jeff Brodsky, John Christianson, Jill Johnson, Paul LaPointe, and Liz Miller for their helpful suggestions and discussions throughout this work and Lars Ellgaard for critical reading of the manuscript.
Funding
A.J.M. acknowledges the support of NIH R15 AREA Grant GM087654 and internal faculty grants from Bennington College.
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Kudze, T., Mendez-Dorantes, C., Jalloh, C.S. et al. Evidence for interaction between Hsp90 and the ER membrane complex. Cell Stress and Chaperones 23, 1101–1115 (2018). https://doi.org/10.1007/s12192-018-0908-z
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DOI: https://doi.org/10.1007/s12192-018-0908-z