HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 47, 48836-48845, November 19, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/47/48836    most recent M407601200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Walerych, D. Articles by Zylicz, M. Search for Related Content PubMed PubMed Citation Articles by Walerych, D. Articles by Zylicz, M. Social Bookmarking                      What's this?

Hsp90 Chaperones Wild-type p53 Tumor Suppressor Protein^*

Dawid Walerych, Grzegorz Kudla, Malgorzata Gutkowska¶, Bartosz Wawrzynow, Lin Muller||, Frank W. King**, Aleksandra Helwak**, Joanna Boros, Alicja Zylicz, and Maciej Zylicz

From the International Institute of Molecular and Cell Biology in Warsaw, 02-109 Warsaw, Poland, the ^¶Nencki Institute of Experimental Biology, Polish Academy of Sciences, 02-093 Warsaw, Poland, the ^||Department Chemie, Technische Universitat Munchen, 85-747 Garching, Germany, and the ^**Institute of Biochemistry and Biophysics, 02-106 Warsaw, Poland

Immortalized human fibroblasts were used to investigate the putative interactions of the Hsp90 molecular chaperone with the wild-type p53 tumor suppressor protein. We show that geldanamycin or radicicol, specific inhibitors of Hsp90, diminish specific wild-type p53 binding to the p21 promoter sequence. Consequently, these inhibitors decrease p21 mRNA levels, which lead to a reduction in cellular p21/Waf1 protein, known to induce cell cycle arrest. In control experiments, we show that neither geldanamycin nor radicicol affect p53 mRNA levels. A minor decrease in p53 protein level following the treatment of human fibroblasts with the inhibitors suggests the potential involvement of Hsp90 in the stabilization of wild-type p53. To support our in vivo findings, we used a reconstituted system with highly purified recombinant proteins to examine the effects of Hsp90 on wild-type p53 binding to the p21 promoter sequence. The human recombinant Hsp90 -isoform as well as bovine brain Hsp90 were purified to homogeneity. Both of these molecular chaperones displayed ATPase activity and the ability to refold heat-inactivated luciferase in a geldanamycin- and radicicol-sensitive manner, suggesting that post-translational modifications are not involved in the modulation of Hsp90 activity. We show that the incubation of recombinant p53 at 37 °C decreases the level of its wild-type conformation and strongly inhibits the in vitro binding of p53 to the p21 promoter sequence. Interestingly, Hsp90 in an ATP-dependent manner can positively modulate p53 DNA binding after incubation at physiological temperature of 37 °C. Other recombinant human chaperones from Hsp70 and Hsp40 families were not able to efficiently substitute Hsp90 in this reaction. Consistent with our in vivo results, geldanamycin can suppress Hsp90 ability to regulate in vitro p53 DNA binding to the promoter sequence. In summary, the results presented in this article state that chaperone activity of Hsp90 is important for the transcriptional activity of genotypically wild-type p53.

Received for publication, July 7, 2004 , and in revised form, September 8, 2004.

^* This work was supported by State Committee for Scientific Research Grant 3P04B02122 (to A. Z.), the Foundation for Polish Science (to M. Z.), V Framework Grant QLRT-2001-02833, and grants from the Deutsche Forschungsgemeinschaft (to J. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

These authors contributed equally to this work and are recipients of a scholarship from the Postgraduate School of Molecular Medicine affiliated with the Medical University of Warsaw.

Present address: Cancer Research Institute, University of California San Francisco, CA.

To whom correspondence should be addressed: International Institute of Molecular and Cell Biology in Warsaw, Trojdena 4 St., 02-109 Warsaw, Poland. Tel.: 48-22-668-50-86; Fax: 48-22-668-50-57; E-mail: zylicz{at}iimcb.gov.pl.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				Y. Fukuyo, M. Inoue, T. Nakajima, R. Higashikubo, N. T. Horikoshi, C. Hunt, A. Usheva, M. L. Freeman, and N. Horikoshi Oxidative Stress Plays a Critical Role in Inactivating Mutant BRAF by Geldanamycin Derivatives Cancer Res., August 1, 2008; 68(15): 6324 - 6330. [Abstract] [Full Text] [PDF]

				S. H. Fisher and L. V. Wray Jr. Bacillus subtilis glutamine synthetase regulates its own synthesis by acting as a chaperone to stabilize GlnR-DNA complexes PNAS, January 22, 2008; 105(3): 1014 - 1019. [Abstract] [Full Text] [PDF]

				O. A. Toogun, D. C. DeZwaan, and B. C. Freeman The Hsp90 Molecular Chaperone Modulates Multiple Telomerase Activities Mol. Cell. Biol., January 1, 2008; 28(1): 457 - 467. [Abstract] [Full Text] [PDF]

				B. Wawrzynow, A. Zylicz, M. Wallace, T. Hupp, and M. Zylicz MDM2 Chaperones the p53 Tumor Suppressor J. Biol. Chem., November 9, 2007; 282(45): 32603 - 32612. [Abstract] [Full Text] [PDF]

				V. Tripathi, A. Ali, R. Bhat, and U. Pati CHIP Chaperones Wild Type p53 Tumor Suppressor Protein J. Biol. Chem., September 28, 2007; 282(39): 28441 - 28454. [Abstract] [Full Text] [PDF]

				M. Sasaki, L. Nie, and C. G. Maki MDM2 Binding Induces a Conformational Change in p53 That Is Opposed by Heat-shock Protein 90 and Precedes p53 Proteasomal Degradation J. Biol. Chem., May 11, 2007; 282(19): 14626 - 14634. [Abstract] [Full Text] [PDF]

				L. T. Gooljarsingh, C. Fernandes, K. Yan, H. Zhang, M. Grooms, K. Johanson, R. H. Sinnamon, R. B. Kirkpatrick, J. Kerrigan, T. Lewis, et al. A biochemical rationale for the anticancer effects of Hsp90 inhibitors: Slow, tight binding inhibition by geldanamycin and its analogues PNAS, May 16, 2006; 103(20): 7625 - 7630. [Abstract] [Full Text] [PDF]

				C. Esser, M. Scheffner, and J. Hohfeld The Chaperone-associated Ubiquitin Ligase CHIP Is Able to Target p53 for Proteasomal Degradation J. Biol. Chem., July 22, 2005; 280(29): 27443 - 27448. [Abstract] [Full Text] [PDF]

				P. Kramata, Y.-P. Lu, Y.-R. Lou, R. N. Singh, S. M. Kwon, and A. H. Conney Patches of Mutant p53-Immunoreactive Epidermal Cells Induced by Chronic UVB Irradiation Harbor the Same p53 Mutations as Squamous Cell Carcinomas in the Skin of Hairless SKH-1 Mice Cancer Res., May 1, 2005; 65(9): 3577 - 3585. [Abstract] [Full Text] [PDF]

				P. Muller, P. Ceskova, and B. Vojtesek Hsp90 Is Essential for Restoring Cellular Functions of Temperature-sensitive p53 Mutant Protein but Not for Stabilization and Activation of Wild-type p53: IMPLICATIONS FOR CANCER THERAPY J. Biol. Chem., February 25, 2005; 280(8): 6682 - 6691. [Abstract] [Full Text] [PDF]

				L. Muller, A. Schaupp, D. Walerych, H. Wegele, and J. Buchner Hsp90 Regulates the Activity of Wild Type p53 under Physiological and Elevated Temperatures J. Biol. Chem., November 19, 2004; 279(47): 48846 - 48854. [Abstract] [Full Text] [PDF]