Hsp90 cleavage by an oxidative stress leads to its client proteins degradation and cancer cell death

Beck, Raphael; Verrax, Julien; Gonze, Thomas; Zappone, Marianne; Pedrosa, Rozangela Curi; Taper, Henryk; Feron, Olivier; Calderon, Pedro Buc

Abstract

The heat shock protein 90 (Hsp90) plays a crucial role in the stability of several proteins that are essential for malignant trans formation. Hsp90 is therefore an interesting therapeutic target for cancer therapy. in this paper, we investigated whether an oxidative stress generated during ascorbate-driven menadione redox cycling (ascorbate/menadione), affects Hsp90 leading to the degradation of some critical proteins and cell death. Unlike 17-AAG, which inhibits Hsp90 but enhances Hsp70 levels, ascorbate/menadione-treated cells present an additional Hsp90 protein band of about 70 kDa as shown by Western blot analysis, suggesting Hsp90 cleavage. This Hsp90 cleavage seems to be a selective phenomenon since it was observed in a large panel of cancer cell lines but not in non-transformed cells. Antibodies raised against either the N-terminus or the C-terminus domains of Hsp90 suggest that the site of cleavage should be located at its N-terminal part. Furthermore, antibodies raised against either the alpha- or the beta-Hsp90 isoform show that Hsp90 beta is cleaved while the a isoform. is down-regulated. We have further shown that different Hsp90 client proteins like Bcr-Abl (a chimerical protein expressed in K562 leukemia cells), RIP and Akt, were degraded when K562 cells were exposed to an oxidative stress. Both Hsp90 cleavage and Bcr-Abl degradation were observed by incubating K562 cells with another H2O2-generating system (glucose/glucose oxidase) and by incubating KU812 cells (another leukemia cell line) with ascorbate/menadione. Due to the major role of Hsp90 in stabilizing oncogenic and mutated proteins, these results may have potential clinical applications. (C) 2008 Elsevier Inc. All rights reserved.

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Título según WOS: ID WOS:000262935900008 Not found in local WOS DB
Título de la Revista: BIOCHEMICAL PHARMACOLOGY
Volumen: 77
Número: 3
Editorial: PERGAMON-ELSEVIER SCIENCE LTD
Fecha de publicación: 2009
Página de inicio: 375
Página final: 383
DOI:

10.1016/j.bcp.2008.10.019

Notas: ISI