Regulation of Molecular Chaperone Gene Transcription Involves the Serine Phosphorylation, 14-3-3ε Binding, and Cytoplasmic Sequestration of Heat Shock Factor 1

Abstract

Heat shock factor 1 (HSF1) regulates the transcription of molecular chaperone hsp genes. However, the cellular control mechanisms that regulate HSF1 activity are not well understood. In this study, we have demonstrated for the first time that human HSF1 binds to the essential cell signaling protein 14-3-3ε. Binding of HSF1 to 14-3-3ε occurs in cells in which extracellular signal regulated kinase (ERK) is activated and blockade of the ERK pathway by treatment with the specific ERK pathway inhibitor PD98059 in vivo strongly suppresses the binding. We previously showed that ERK1 phosphorylates HSF1 on serine 307 and leads to secondary phosphorylation by glycogen synthase kinase 3 (GSK3) on serine 303 within the regulatory domain and that these phosphorylation events repress HSF1. We show here that HSF1 binding to 14-3-3ε requires HSF1 phosphorylation on serines 303 and 307. Furthermore, the serine phosphorylation-dependent binding of HSF1 to 14-3-3ε results in the transcriptional repression of HSF1 and its sequestration in the cytoplasm. Leptomycin B, a specific inhibitor of nuclear export receptor CRM1, was found to reverse the cytoplasmic sequestration of HSF1 mediated by 14-3-3ε, suggesting that CRM1/14-3-3ε directed nuclear export plays a major role in repression of HSF1 by the ERK/GSK3/14-3-3ε pathway. Our experiments indicate a novel pathway for HSF1 regulation and suggest a mechanism for suppression of its activity during cellular proliferation.

ACKNOWLEDGMENTS

We thank Michael Yaffe for the gift of 14-3-3ε-GST plasmid, Brendan Price for help and advice, and Sorab N. Dalal and James DeCaprio for HA-tagged 14-3-3ε expression vector. We also appreciate helpful suggestions and comments from our colleagues, Yue Xie, Dan Tang, and Alexzander Asea during preparation of the manuscript.

This work was supported by National Institutes of Health grants CA47407, CA31303, CA50642, and CA77465.