The ability to infect cells latently helps HIV to establish persistent infections despite strong humoral and cellular immune responses against the viral proteins and remains the single greatest obstacle to the development of therapies that result in the elimination of HIV from infected individuals. This proposal builds on some remarkable observations we have recently made demonstrating that the activation of HIV transcription is an extraordinarily dynamic process. We have found using high resolution chromatin immunoprecipitation (ChIP) experiments that the recruitment of RNA polymerase; TFIIH and numerous other components of the transcription machinery to the proviral promoter rapidly rises and falls as NF-kappaB enters and exits the nucleus. The discovery that the transcriptional responses of HIV to cellular activation signals is intermittent, and not simply an irreversible response to an initial signaling event, was completely unexpected. In complementary studies, we also plan to investigate the molecular mechanisms utilized by HIV to enter latency. We have recently found that transcription from integrated lentiviral vectors that carry Tat in cis progressively diminishes over a period of 4 to 6 weeks until the virus enters a latent state. The shutdown in transcription appears to be associated with a loss of Tat activity since mutations in Tat that reduce its activity lead to more rapid viral shutdown. These observations lead to the hypothesis that HIV enters a latent state once initiation rates have fallen to a level where threshold levels of Tat are no longer produced. Activation of NF-kappaB and related transcription factors restores transcription initiation and re-establishes the Tat feedback mechanism leading to high levels of viral RNA synthesis. Because reactivation of HIV transcription in response to cellular stimulation is a highly dynamic process in which multiple transcription factors are rapidly recruited and exchanged at the promoter, multiple signaling pathways can be used to regulate both the timing and magnitude of the transcriptional response of HIV to external stimuli. We plan to rigorously test this hypothesis using a combination of biochemical studies of HIV promoter activation in model cellular systems, and the development of primary cell models for proviral quiescence and latency. These studies promise to lead to a detailed new understanding of the specific factors that drive HIV latency and suggest new avenues for antiviral research. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI067093-01A1
Application #
7230319
Study Section
Special Emphasis Panel (ZRG1-AARR-A (03))
Program Officer
Young, Janet M
Project Start
2007-02-01
Project End
2012-01-31
Budget Start
2007-02-01
Budget End
2008-01-31
Support Year
1
Fiscal Year
2007
Total Cost
$386,250
Indirect Cost
Name
Case Western Reserve University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
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Das, Biswajit; Dobrowolski, Curtis; Shahir, Abdel-Malek et al. (2015) Short chain fatty acids potently induce latent HIV-1 in T-cells by activating P-TEFb and multiple histone modifications. Virology 474:65-81
Mbonye, Uri R; Wang, Benlian; Gokulrangan, Giridharan et al. (2015) Phosphorylation of HEXIM1 at Tyr271 and Tyr274 Promotes Release of P-TEFb from the 7SK snRNP Complex and Enhances Proviral HIV Gene Expression. Proteomics 15:2078-86
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Mbonye, Uri; Karn, Jonathan (2014) Transcriptional control of HIV latency: cellular signaling pathways, epigenetics, happenstance and the hope for a cure. Virology 454-455:328-39
Jadlowsky, Julie K; Wong, Julian Y; Graham, Amy C et al. (2014) Negative elongation factor is required for the maintenance of proviral latency but does not induce promoter-proximal pausing of RNA polymerase II on the HIV long terminal repeat. Mol Cell Biol 34:1911-28
Friedman, Julia; Cho, Won-Kyung; Chu, Chung K et al. (2011) Epigenetic silencing of HIV-1 by the histone H3 lysine 27 methyltransferase enhancer of Zeste 2. J Virol 85:9078-89
Kim, Young Kyeung; Mbonye, Uri; Hokello, Joseph et al. (2011) T-cell receptor signaling enhances transcriptional elongation from latent HIV proviruses by activating P-TEFb through an ERK-dependent pathway. J Mol Biol 410:896-916
Karn, Jonathan (2011) The molecular biology of HIV latency: breaking and restoring the Tat-dependent transcriptional circuit. Curr Opin HIV AIDS 6:4-11
Mbonye, Uri; Karn, Jonathan (2011) Control of HIV latency by epigenetic and non-epigenetic mechanisms. Curr HIV Res 9:554-67

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