HIV-1 infection depends on a series of molecular events involving biochemical interactions between viral components and host factors, resulting in fusion of viral and host membranes, reverse transcription, uncoating, nuclear entry, and integration. A key knowledge gap in the early stages of infection is the role of the viral capsid i facilitating reverse transcription. Mutations in the CA protein that destabilize the viral capsid result in impaired reverse transcription, indicating that the integrity of the viral capsid is critcal for efficient viral DNA synthesis. HIV-1 reverse transcriptase is a low processivity enzyme and dissociates from the template during strand transfer steps of reverse transcription. Employing a biochemical approach involving purified HIV-1 cores, we will test the hypothesis that the viral capsid serves as a vessel to maintain the local concentration of RT during reverse transcription and will study the role of restrictive host factors in HIV-1 capsid recognition and uncoating. We will also identify novel host factors that bind the HIV-1 capsid. The work will be organized by the following Specific Aims:
Aim 1. To determine the mechanism by which the viral capsid facilitates HIV-1 reverse transcription.
Aim 2. Identification of intersubunit interfaces critical for HIV-1 uncoating.
Aim 3. To determine the effect of CPSF6-358 on HIV-1 uncoating.
Aim 4. To determine the stoichiometric requirement for functional capsid recognition by TRIM5 restriction factors.
Aim 5. Identification of novel capsid-interacting host factors. These studies will answer fundamental question in retrovirus biology and will clarify the mechanisms of restriction by endogenous host cell factors.

Public Health Relevance

HIV/AIDS remains a global epidemic and public health threat. Despite the development of effective antiretroviral drugs, therapy is not curative and is therefore life-long. A detailed understanding of the uncoating step in HIV-1 infection will lead to the development of an improved therapeutic arsenal and novel insights into interactions between the virus and its cellular targets.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI076121-05A1
Application #
8467218
Study Section
AIDS Molecular and Cellular Biology Study Section (AMCB)
Program Officer
Sanders, Brigitte E
Project Start
2008-03-15
Project End
2017-02-28
Budget Start
2013-03-20
Budget End
2014-02-28
Support Year
5
Fiscal Year
2013
Total Cost
$366,600
Indirect Cost
$131,600
Name
Vanderbilt University Medical Center
Department
Pathology
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
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Shi, Jiong; Friedman, David B; Aiken, Christopher (2013) Retrovirus restriction by TRIM5 proteins requires recognition of only a small fraction of viral capsid subunits. J Virol 87:9271-8
Yufenyuy, Ernest L; Aiken, Christopher (2013) The NTD-CTD intersubunit interface plays a critical role in assembly and stabilization of the HIV-1 capsid. Retrovirology 10:29
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Roa, Amanda; Hayashi, Fumiaki; Yang, Yang et al. (2012) RING domain mutations uncouple TRIM5? restriction of HIV-1 from inhibition of reverse transcription and acceleration of uncoating. J Virol 86:1717-27
Yang, Ruifeng; Shi, Jiong; Byeon, In-Ja L et al. (2012) Second-site suppressors of HIV-1 capsid mutations: restoration of intracellular activities without correction of intrinsic capsid stability defects. Retrovirology 9:30
Du, Shoucheng; Betts, Laurie; Yang, Ruifeng et al. (2011) Structure of the HIV-1 full-length capsid protein in a conformationally trapped unassembled state induced by small-molecule binding. J Mol Biol 406:371-86
Shi, Jiong; Zhou, Jing; Shah, Vaibhav B et al. (2011) Small-molecule inhibition of human immunodeficiency virus type 1 infection by virus capsid destabilization. J Virol 85:542-9
Shah, Vaibhav B; Aiken, Christopher (2011) In vitro uncoating of HIV-1 cores. J Vis Exp :

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