Abstract
Heat shock protein 90 (Hsp90) is an essential molecular chaperone in eukaryotes, as it regulates diverse signal transduction nodes that integrate numerous environmental cues to maintain cellular homeostasis. Hsp90 also is secreted from normal and transformed cells and regulates cell motility. Here, we have identified a conserved hydrophobic motif in a β-strand at the boundary between the N domain and charged linker of Hsp90, whose mutation not only abrogated Hsp90 secretion but also inhibited its function. These Hsp90 mutants lacked chaperone activity in vitro and failed to support yeast viability. Notably, truncation of the charged linker reduced solvent accessibility of this β-strand and restored chaperone activity to these mutants. These data underscore the importance of β-strand 8 for Hsp90 function and demonstrate that the functional consequences of weakened hydrophobic contacts in this region are reversed by charged-linker truncation.
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Acknowledgements
We are indebted to K. Beebe for comments on the manuscript. This work is supported by US National Cancer Institute intramural research funds. S.T. is partially supported by a fellowship from the Japanese Society of the Promotion of Science.
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S.T. and M.M. performed most of the experiments and helped interpret the data, design experiments and write the manuscript; C.G., B.T.S., C.-T.L., W.X., L.H., M.H. and A.A.K. performed some of the experiments; B.P., J.B. and M.P.M. designed some of the experiments and interpreted the data collected; M.P.M. and J.B. also contributed to writing part of the manuscript; J.B.T. contributed to the design and interpretation of some of the experiments; C.P. designed and performed some of the experiments, interpreted the data collected and contributed to writing part of the manuscript; L.N. oversaw the entire project, designed experiments, interpreted the data and wrote the paper.
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Tsutsumi, S., Mollapour, M., Graf, C. et al. Hsp90 charged-linker truncation reverses the functional consequences of weakened hydrophobic contacts in the N domain. Nat Struct Mol Biol 16, 1141–1147 (2009). https://doi.org/10.1038/nsmb.1682
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DOI: https://doi.org/10.1038/nsmb.1682
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