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J. Biol. Chem., Vol. 276, Issue 48, 44633-44640, November 30, 2001
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From the Tat stimulates human immunodeficiency virus, type
1 (HIV-1), transcription elongation by recruitment of the human
transcription elongation factor P-TEFb, consisting of CDK9 and cyclin
T1, to the TAR RNA structure. It has been demonstrated further that
CDK9 phosphorylation is required for high affinity binding of
Tat/P-TEFb to the TAR RNA structure and that the state of P-TEFb
phosphorylation may regulate Tat transactivation. We now demonstrate
that CDK9 phosphorylation is uniquely regulated in the HIV-1
preinitiation and elongation complexes. The presence of TFIIH in the
HIV-1 preinitiation complex inhibits CDK9 phosphorylation. As TFIIH is
released from the elongation complex between +14 and +36, CDK9
phosphorylation is observed. In contrast to the activity in the
"soluble" complex, phosphorylation of CDK9 is increased by the
presence of Tat in the transcription complexes. Consistent with these
observations, we have demonstrated that purified TFIIH directly
inhibits CDK9 autophosphorylation. By using recombinant TFIIH
subcomplexes, our results suggest that the XPB subunit of TFIIH is
responsible for this inhibition of CDK9 phosphorylation. Interestingly,
our results further suggest that the phosphorylated form of CDK9 is the
active kinase for RNA polymerase II carboxyl-terminal domain phosphorylation.
TFIIH Inhibits CDK9 Phosphorylation during Human Immunodeficiency
Virus Type 1 Transcription*
§,
,, and§§
Virus Tumor Biology Section, Basic Research
Laboratory, Division of Basic Sciences, NCI, National Institutes of
Health, Bethesda, Maryland 20892, the § Department of
Biochemistry and Molecular Biology, George Washington University
Medical Center, Washington, D. C. 20037, the Laboratory of
Molecular Embryology, NICHD, Bethesda, Maryland 20892, the
** Department of Biochemistry, University of Iowa,
Iowa City, Iowa 52242, and the Institut
de Genetique et de Biologie Moleculaire et Cellulaire, CNRS/INSERM/ULP,
B. P. 163, 67404 Illkirch Cedex, C. U. de
Strasbourg, France
*
This work was supported in part by the Intramural AIDS
Targeted Antiviral Program from the Office of the Director, National Institutes of Health (to J. N. B.).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.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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