Coordinated Conformational Processing of the Tumor Suppressor Protein p53 by the Hsp70 and Hsp90 Chaperone Machineries

p53, the guardian of the genome, requires chaperoning by Hsp70 and Hsp90. However, how the two chaperone machineries affect p53 conformation and regulate its function remains elusive. We found that Hsp70, together with Hsp40, unfolds p53 in an ATP-dependent reaction. This unfolded state of p53 is susceptible to aggregation after release induced by the nucleotide exchange factor Bag-1. However, when Hsp90 and the adaptor protein Hop are present, p53 is transferred from Hsp70 to Hsp90, allowing restoration of the native state upon ATP hydrolysis. Our results suggest that the p53 conformation is constantly remodeled by the two major chaperone machineries. This connects p53 activity to stress, and the levels of free molecular chaperones are important factors regulating p53 activity. Together, our findings reveal an intricate interplay and cooperation of Hsp70 and Hsp90 in regulating the conformation of a client. [Display omitted] •p53 requires Hsp70 and Hsp90 for regulating its DNA binding activity•Hsp70 inactivates p53 at physiological temperatures by unfolding it•Hsp90 folds p53 to native conformation in an ATP-dependent process•p53 conformation constantly switches between Hsp70- and Hsp90-bound states p53 activity is tightly regulated by opposing effects of Hsp70 and Hsp90 on its conformation. Hsp70-Hsp40 unfolds p53, whereas Hsp90 and Hop fold it into native conformation upon ATP hydrolysis. These findings establish a concept of how DNA binding of transcription factors can be tightly regulated by Hsp70 and Hsp90.