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Journal of Virology, December 2003, p. 13028-13035, Vol. 77, No. 24
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.24.13028-13035.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The Central Region of Human T-Cell Leukemia Virus Type 1 Tax Protein Contains Distinct Domains Involved in Subunit Dimerization

Jihane Basbous,1,2 Ali Bazarbachi,2 Claude Granier,3 Christian Devaux,1 and Jean-Michel Mesnard1*

Laboratoire Infections Rétrovirales et Signalisation Cellulaire, CNRS/UM I UMR 5121/IFR 122, Institut de Biologie, 34960 Cedex 2, Montpellier,1 CNRS/UM I UMR 9921/IFR 122, Faculté de Pharmacie, 34060 Montpellier,France,3 Departments of Internal Medicine and Biochemistry, American University of Beirut, Beirut, Lebanon2

Received 12 June 2003/ Accepted 5 September 2003

The Tax protein of human T-cell leukemia virus type 1 (HTLV-1) can form homodimers. Tax dimerization contributes to optimal Tax activity involved in transactivation of the HTLV-1 promoter. The mechanisms used to form specific Tax dimers are poorly understood because the domains that mediate such interactions have not been clearly characterized. Here we have used different approaches (the two-hybrid assay in yeast, the glutathione S-transferase pull-down assay, and the Spot method) to study Tax-Tax interactions. Our results indicate that the integrity of the sequence of Tax, except for the last 16 amino acids (residues 338 to 353), is critical, suggesting that Tax dimerization is dictated more by secondary structure than by primary structure. We were, however, able to delimit a central region involved in Tax self-association that encompasses the residues 127 to 228. This region can be divided into three subdomains of dimerization: DD1 (residues 127 to 146), DD2 (residues 181 to 194), and DD3 (residues 213 to 228). Moreover, the Tax mutants M22 (T130A and L131S) and M29 (K189A and R190S), with amino acid substitutions located in DD1 and DD2, respectively, were found to be impaired in Tax self-association.

* Corresponding author. Mailing address: Laboratoire Infections Rétrovirales et Signalisation Cellulaire, Institut de Biologie, 4 Bd. Henri IV, CS 89508, 34960 Cedex 2, Montpellier, France. Phone: (33) 4-67-60-86-60. Fax: (33) 4-67-60-44-20. E-mail: jean-michel.mesnard{at}univ-montp1.fr.

Journal of Virology, December 2003, p. 13028-13035, Vol. 77, No. 24
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.24.13028-13035.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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