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Originally published In Press as doi:10.1074/jbc.M007901200 on January 18, 2001

J. Biol. Chem., Vol. 276, Issue 16, 12959-12966, April 20, 2001
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HIV-1 Tat Protein Interacts with Mammalian Capping Enzyme and Stimulates Capping of TAR RNA*

Ya-Lin ChiuDagger , Elizabeth CoronelDagger , C. Kiong Ho§, Stewart Shuman§, and Tariq M. RanaDagger

From the Dagger  Department of Pharmacology, UMDNJ-Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, and the § Molecular Biology Program, Sloan-Kettering Institute, New York, New York 10021

HIV gene expression is subject to a transcriptional checkpoint, whereby negative transcription elongation factors induce an elongation block that is overcome by HIV Tat protein in conjunction with P-TEFb. P-TEFb is a cyclin-dependent kinase that catalyzes Tat-dependent phosphorylation of Ser-5 of the Pol II C-terminal domain (CTD). Ser-5 phosphorylation confers on the CTD the ability to recruit the mammalian mRNA capping enzyme (Mce1) and stimulate its guanylyltransferase activity. Here we show that Tat spearheads a second and novel pathway of capping enzyme recruitment and activation via a direct physical interaction between the C-terminal domain of Tat and Mce1. Tat stimulates the guanylyltransferase and triphosphatase activities of Mce1 and thereby enhances the otherwise low efficiency of cap formation on a TAR stem-loop RNA. Our findings suggest that multiple mechanisms exist for coupling transcription elongation and mRNA processing.

* This work was supported by National Institutes of Health Grants AI43198 (to T. M. R.) and GM52470 (to S. S.).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: UMDNJ-Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, NJ 08854. Tel.: 732-235-4082; Fax: 732-235-3235; E-mail: rana@umdnj.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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