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Originally published In Press as doi:10.1074/jbc.M006804200 on August 15, 2000

J. Biol. Chem., Vol. 275, Issue 44, 34314-34319, November 3, 2000
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HIV-1 TAR RNA Enhances the Interaction between Tat and Cyclin T1*

Jing ZhangDagger , Natarajan TamilarasuDagger , Seongwoo HwangDagger , Mitchell E. Garber§, Ikramul HuqDagger , Katherine A. Jones§, and Tariq M. RanaDagger

From the Dagger  Department of Pharmacology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway, New Jersey 08854 and the § Regulatory Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037

Human immunodeficiency virus, type 1 (HIV-1), Tat activates elongation of RNA polymerase II transcription at the HIV-1 promoter through interaction with the cyclin T1 (CycT1) subunit of the positive transcription elongation factor complex, P-TEFb. Binding of Tat to CycT1 induces cooperative binding of the P-TEFb complex onto nascent HIV-1 TAR RNA. Here the specific interaction between Tat protein, human cyclin T1, and HIV-1 TAR RNA was analyzed by fluorescence resonance energy transfer, using fluorescein-labeled TAR RNA and a rhodamine-labeled Tat protein synthesized through solid-phase chemistry. We find that CycT1 remodels the structure of Tat to enhance its affinity for TAR RNA and that TAR RNA further enhances the interaction between Tat and CycT1. We conclude that TAR RNA nucleates the formation of the Tat·P-TEFb complex through an induced fit mechanism.

* This work was supported in part by National Institutes of Health Grant AI 41404 (to T. M. R.).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.

Recipient of a Research Career Development award from the National Institutes of Health. To whom correspondence should be addressed: Dept. of Pharmacology, UMDNJ-Robert Wood Johnson Medical School, 675 Hoes Ln., Piscataway, NJ 08854. Tel.: 732-235-4082; Fax: 732- 235-3235; E-mail: rana@umdnj.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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