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J. Biol. Chem., Vol. 279, Issue 1, 495-508, January 2, 2004

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Protein Profile of Tax-associated Complexes^*

Kaili Wu, Maria Elena Bottazzi, Cynthia de la Fuente, Longwen Deng, Scott D. Gitlin¶, Anil Maddukuri, Shabnam Dadgar, Hong Li||, Akos Vertes**, Anne Pumfery, and Fatah Kashanchi

From the Departments of Biochemistry and Molecular Biology and Microbiology and Tropical Medicine, School of Medicine and Health Sciences, The George Washington University, Washington, D. C. 20037, ^||Department of Biochemistry and Molecular Biology, University of Medicine and Dentistry, New Jersey Medical School, Newark, New Jersey 07103, ^¶Department of Internal Medicine, Division of Hematology/Oncology, University of Michigan, Ann Arbor, Michigan 48109-0640, and ^**Department of Chemistry, The George Washington University, Washington, D. C. 20052

Infection with human T-cell leukemia virus type 1 (HTLV-1) results in adult T-cell leukemia and HTLV-1-associated myelopathy/tropical spastic paraparesis. Tax, a 40-kDa protein, regulates viral and cellular transcription, host signal transduction, the cell cycle, and apoptosis. Tax has been shown to modulate cellular CREB and NFB pathways; however, to date, its role in binding to various host cellular proteins involved in tumorigenesis has not been fully described. In this study, we describe the Tax-associated proteins and their functions in cells using several approaches. Tax eluted from a sizing column mostly at an apparent molecular mass of 1800 kDa. Following Tax immunoprecipitation, washes with high salt buffer, two-dimensional gel separation, and mass spectrometric analysis, a total of 32 proteins was identified. Many of these proteins belong to the signal transduction and cytoskeleton pathways and transcription/chromatin remodeling. A few of these proteins, including TXBP151, have been shown previously to bind to Tax. The interaction of Tax with small GTPase-cytoskeleton proteins, such as ras GAP^1m, Rac1, Cdc42, RhoA, and gelsolin, indicates how Tax may regulate migration, invasion, and adhesion in T-cell cancers. Finally, the physical and functional association of Tax with the chromatin remodeling SWI/SNF complex was assessed using in vitro chromatin remodeling assays, chromatin remodeling factor BRG1 mutant cells, and RNA interference experiments. Collectively, Tax is able to bind and regulate many cellular proteins that regulate transcription and cytoskeletal related pathways, which might explain the pleiotropic effects of Tax leading to T-cell transformation and leukemia in HTLV-1-infected patients.

Received for publication, September 10, 2003

^* This work was supported in part by National Institutes of Health Grants AI44357, AI43894, and 13969 (to F. K.), by the Alexandrine and Alexander Sinsheimer Foundation (to K. W., H. L., and L. D.), and by Research Enhancement Fund 111501 from The George Washington University (to A. V. and F. K.). 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.

To whom correspondence should be addressed: Dept. of Biochemistry and Molecular Biology, School of Medicine, Ross Hall, Rm. 552, The George Washington University, 2300 Eye St., NW, Washington, D. C. 20037. Tel.: 202-994-1782; Fax: 202-994-1780; E-mail: bcmfxk{at}gwumc.edu.

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