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Journal of Virology, April 2005, p. 4396-4406, Vol. 79, No. 7
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.7.4396-4406.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

TFII-I Regulates Induction of Chromosomally Integrated Human Immunodeficiency Virus Type 1 Long Terminal Repeat in Cooperation with USF

Jiguo Chen,1,{dagger} Tom Malcolm,1,{dagger} Mario C. Estable,2,3 Robert G. Roeder,3 and Ivan Sadowski1*

Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia,1 Department of Chemistry and Biology, Ryerson University, Toronto, Ontario, Canada,2 Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, New York, New York3

Received 25 September 2004/ Accepted 11 November 2004

Human immunodeficiency virus type 1 (HIV-1) replication is coupled to T-cell activation through its dependence on host cell transcription factors. Despite the enormous sequence variability of these factors, several cis elements for host factors are highly conserved within the 5' long terminal repeats (LTRs) of viruses from AIDS patients; among these is the RBEIII upstream element for the Ras response element binding factor 2 (RBF-2). Here we show that RBF-2 is comprised of a USF1/USF2 heterodimer and TFII-I, which bind cooperatively to RBEIII. Recombinant USF1/USF2 binds to the RBEIII core sequence 160-fold less efficiently than it binds to an E box element, but the interaction with RBEIII is stimulated by TFII-I. Chromosomally integrated HIV-1 LTRs bearing an RBEIII mutation have slightly elevated basal transcription in unstimulated Jurkat cells but are unresponsive to cross-linking of the T-cell receptor or stimulation with phorbol myristate acetate (PMA) and ionomycin. Induction is inhibited by dominant interfering USF and TFII-I but not by the dominant negative I-{kappa}B protein. USF1, USF2, and TFII-I bind to the integrated wild-type LTR in unstimulated cells and become phosphorylated during the induction of transcription upon stimulation with PMA. These results demonstrate that USF1/USF2 and TFII-I interact cooperatively at the upstream RBEIII element and are necessary for the induction of latent HIV-1 in response to T-cell activation signals.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, UBC, 2146 Health Sciences Mall, Vancouver, BC, V6T 1Z3, Canada. Phone: (604) 822-4524. Fax: (604) 822-9311. E-mail: sadowski{at}interchange.ubc.ca.

{dagger} J.C. and T.M. contributed equally to this research.

Journal of Virology, April 2005, p. 4396-4406, Vol. 79, No. 7
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.7.4396-4406.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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