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Title: Identification and functional analysis of a second RBF-2 binding site within the HIV-1 promoter

Abstract

Transcription from the HIV-1 long terminal repeat (LTR) is mediated by numerous host transcription factors. In this study we characterized an E-box motif (RBE1) within the core promoter that was previously implicated in both transcriptional activation and repression. We show that RBE1 is a binding site for the RBF-2 transcription factor complex (USF1, USF2, and TFII-I), previously shown to bind an upstream viral element, RBE3. The RBE1 and RBE3 elements formed complexes of identical mobility and protein constituents in gel shift assays, both with Jurkat T-cell nuclear extracts and recombinant USF/TFII-I. Furthermore, both elements are regulators of HIV-1 expression; mutations in LTR-luciferase reporters and in HIV-1 molecular clones resulted in decreased transcription, virion production, and proviral expression in infected cells. Collectively, our data indicate that RBE1 is a bona fide RBF-2 binding site and that the RBE1 and RBE3 elements are necessary for mediating proper transcription from the HIV-1 LTR.

Authors:
 [1];  [2];  [1];  [2];  [3];  [3];  [1]
  1. Department of Biochemistry and Molecular Biology, Molecular Epigenetics, Life Sciences Institute, University of British Columbia, 2350 Health Sciences Mall, Vancouver, B.C. V6T 1Z3 (Canada)
  2. Department of Microbiology, Mount Sinai School of Medicine, NY, NY, 10029 (United States)
  3. (United States)
Publication Date:
OSTI Identifier:
21587889
Resource Type:
Journal Article
Journal Name:
Virology
Additional Journal Information:
Journal Volume: 418; Journal Issue: 1; Other Information: DOI: 10.1016/j.virol.2011.07.002; PII: S0042-6822(11)00293-5; Copyright (c) 2011 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0042-6822
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; AIDS VIRUS; FUNCTIONAL ANALYSIS; LUCIFERASE; MUTATIONS; TRANSCRIPTION FACTORS; ENZYMES; MATHEMATICS; MICROORGANISMS; ORGANIC COMPOUNDS; OXIDASES; OXIDOREDUCTASES; PARASITES; PROTEINS; VIRUSES

Citation Formats

Dahabieh, Matthew S., E-mail: dahabieh@interchange.ubc.ca, Ooms, Marcel, E-mail: marcel.ooms@mssm.edu, Malcolm, Tom, E-mail: tmalc1@yahoo.com, Simon, Viviana, E-mail: viviana.simon@mssm.edu, Department of Medicine, Mount Sinai School of Medicine, NY, NY, 10029, The Global Health and Emerging Pathogens Institute, Mount Sinai School of Medicine, NY, NY, 10029, and Sadowski, Ivan, E-mail: ijs.ubc@gmail.com. Identification and functional analysis of a second RBF-2 binding site within the HIV-1 promoter. United States: N. p., 2011. Web. doi:10.1016/j.virol.2011.07.002.
Dahabieh, Matthew S., E-mail: dahabieh@interchange.ubc.ca, Ooms, Marcel, E-mail: marcel.ooms@mssm.edu, Malcolm, Tom, E-mail: tmalc1@yahoo.com, Simon, Viviana, E-mail: viviana.simon@mssm.edu, Department of Medicine, Mount Sinai School of Medicine, NY, NY, 10029, The Global Health and Emerging Pathogens Institute, Mount Sinai School of Medicine, NY, NY, 10029, & Sadowski, Ivan, E-mail: ijs.ubc@gmail.com. Identification and functional analysis of a second RBF-2 binding site within the HIV-1 promoter. United States. doi:10.1016/j.virol.2011.07.002.
Dahabieh, Matthew S., E-mail: dahabieh@interchange.ubc.ca, Ooms, Marcel, E-mail: marcel.ooms@mssm.edu, Malcolm, Tom, E-mail: tmalc1@yahoo.com, Simon, Viviana, E-mail: viviana.simon@mssm.edu, Department of Medicine, Mount Sinai School of Medicine, NY, NY, 10029, The Global Health and Emerging Pathogens Institute, Mount Sinai School of Medicine, NY, NY, 10029, and Sadowski, Ivan, E-mail: ijs.ubc@gmail.com. Thu . "Identification and functional analysis of a second RBF-2 binding site within the HIV-1 promoter". United States. doi:10.1016/j.virol.2011.07.002.
@article{osti_21587889,
title = {Identification and functional analysis of a second RBF-2 binding site within the HIV-1 promoter},
author = {Dahabieh, Matthew S., E-mail: dahabieh@interchange.ubc.ca and Ooms, Marcel, E-mail: marcel.ooms@mssm.edu and Malcolm, Tom, E-mail: tmalc1@yahoo.com and Simon, Viviana, E-mail: viviana.simon@mssm.edu and Department of Medicine, Mount Sinai School of Medicine, NY, NY, 10029 and The Global Health and Emerging Pathogens Institute, Mount Sinai School of Medicine, NY, NY, 10029 and Sadowski, Ivan, E-mail: ijs.ubc@gmail.com},
abstractNote = {Transcription from the HIV-1 long terminal repeat (LTR) is mediated by numerous host transcription factors. In this study we characterized an E-box motif (RBE1) within the core promoter that was previously implicated in both transcriptional activation and repression. We show that RBE1 is a binding site for the RBF-2 transcription factor complex (USF1, USF2, and TFII-I), previously shown to bind an upstream viral element, RBE3. The RBE1 and RBE3 elements formed complexes of identical mobility and protein constituents in gel shift assays, both with Jurkat T-cell nuclear extracts and recombinant USF/TFII-I. Furthermore, both elements are regulators of HIV-1 expression; mutations in LTR-luciferase reporters and in HIV-1 molecular clones resulted in decreased transcription, virion production, and proviral expression in infected cells. Collectively, our data indicate that RBE1 is a bona fide RBF-2 binding site and that the RBE1 and RBE3 elements are necessary for mediating proper transcription from the HIV-1 LTR.},
doi = {10.1016/j.virol.2011.07.002},
journal = {Virology},
issn = {0042-6822},
number = 1,
volume = 418,
place = {United States},
year = {2011},
month = {9}
}