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Originally published In Press as doi:10.1074/jbc.M110233200 on December 17, 2001

J. Biol. Chem., Vol. 277, Issue 9, 7157-7164, March 1, 2002
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HMGB1 and HMGB2 Cell-specifically Down-regulate the p53- and p73-dependent Sequence-specific Transactivation from the Human Bax Gene Promoter*

Michal StrosDagger §, Toshinori Ozaki, Alena BacíkováDagger , Hajime Kageyama, and Akira Nakagawara||

From the Dagger  Institute of Biophysics, Academy of Sciences of the Czech Republic, 612 65 Brno, Czech Republic, and the  Division of Biochemistry, Chiba Cancer Center Research Institute, Chiba 260-8717, Japan

The recently cloned gene p73 is a close homologue of p53, which is a crucial tumor suppressor gene for preventing the malignant transformation of cells by inducing cell cycle arrest and apoptosis. Previous reports have shown that architectural DNA-bending/looping chromosomal proteins HMGB1 and HMGB2 (formerly known as HMG1 and HMG2), which function in a number of biological processes including transcription and DNA repair, interact in vitro with p53 and stimulate p53 binding to DNA containing p53 consensus sites. Here, we report that HMGB1 physically interacts with two splicing variants of p73, alpha  and beta  (pull-down assay), and enhances binding of p73 to specific cognate DNA sites (gel-shift assay). Both HMG box domains of HMGB1, A and B, interact with p73alpha . Association of HMGB1 with p73, like the demonstrated ability of HMGB1 to stimulate p73 binding to different p53-responsive elements, requires the oligomerization region and/or region between DNA-binding domain and oligomerization domain of p73 (residues 312-381). Transient transfections revealed that ectopically expressed or endogenous HMGB1 and HMGB2 (antisense strategy) significantly inhibit in vivo both p73alpha /beta - and p53-dependent transactivation from the Bax gene promoter (and much less from Mdm2 and p21waf1 promoters) in p53-deficient SAOS-2 cells. In contrast, HMGB1 and HGMB2 stimulate p73- or p53-dependent transactivation in p53-deficient H1299 cells, irrespective of the promoter used. Our results suggest that ubiquitously expressed HMGB1 and HMGB2 have potential to cell- and promoter-specifically down- or up-regulate in vivo transcriptional activity of different members of the p53 family. A possible mechanism of HMGB1-mediated modulation of p73- and p53-dependent transactivation is discussed.

* This work was supported in part by Grants A7004902/1999 and IAA5004105 from the Internal Grant Agency of the Academy of Sciences of the Czech Republic and Grants 301/99/0691 and 301/02/0952 from the Grant Agency of the Czech Republic (to M. S.)., and by a grant-in-aid from the Ministry of Health and Welfare of Japan for a New 10-year Strategy of Cancer Control (to A. N.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Supported by the Foundation for Promotion of Cancer Research. To whom correspondence may be addressed: Inst. of Biophysics, Academy of Sciences of the Czech Republic, Královopolská 135, 612 65 Brno, Czech Republic. Tel.: 420-5-41517183; Fax: 420-5-41211293; E-mail: stros@ibp.cz.

|| To whom correspondence may be addressed: Division of Biochemistry, Chiba Cancer Center Research Inst., 666-2 Nitona, Chuoh-ku, Chiba 260-8717, Japan. Tel.: 81-43-264-5431; Fax: 81-43-265-4459; E-mail: akiranak@chiba-ccri.chuo.chiba.jp.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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