The last few years have witnessed a paradigm shift from the view that HIV-1 genetic recombination is a somewhat rare error mechanism to the recognition that it is frequent and is probably an integral part of viral DNA synthesis. This proposal describes a basic molecular genetics study with the long-term objectives of better understanding HIV genetic recombination mechanisms and the factors that modulate it. In the proposed work, a series of modified HIV-1 genomes will be used in single replication cycle tissue culture-based assays to test basic properties of recombination and how properties of RNA templates like those that the virus encounters in vivo affect the frequency and outcomes of recombinogenic template switching.
The specific aims are 1) To develop new assays for studying recombination related properties during HIV-1 replication in cultured cells, including testing predictions of the model that some HIV variability may result from transductive recombination. 2) To test the effects of different physical contexts of potential recombination substrates, such as the nature of dimer linkages, on the substrates abilities to contribute to reverse transcription products, and 3) To examine properties of template switch complexes including the determinants of transfer point, and whether or not mismatch excision and/or extension is apparent among template switch products. The proposed work will advance basic understanding of HIV-1 reverse transcription properties. It is important that this work be performed with HIV-1 because these properties may differ from analogous processes for simple retroviruses. Gamma- and other simple retroviruses' reverse transcription properties during replication have been better studied than have those of HIV. Although it has been assumed that simple retroviruses' properties represent those of HIV, recent advances provide examples of significant differences. Reverse transcription remains the foremost target of antiretroviral drugs, and a more thorough understanding of basic features of the process during HIV replication will be useful to the design and implementation of AIDS antiviral strategies. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM063479-06S1
Application #
7241038
Study Section
AIDS Discovery and Development of Therapeutics Study Section (ADDT)
Program Officer
Portnoy, Matthew
Project Start
2001-04-01
Project End
2008-03-31
Budget Start
2006-04-01
Budget End
2007-03-31
Support Year
6
Fiscal Year
2006
Total Cost
$39,417
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
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Johnson, Silas F; Telesnitsky, Alice (2010) Retroviral RNA dimerization and packaging: the what, how, when, where, and why. PLoS Pathog 6:e1001007
Onafuwa-Nuga, Adewunmi; Telesnitsky, Alice (2009) The remarkable frequency of human immunodeficiency virus type 1 genetic recombination. Microbiol Mol Biol Rev 73:451-80, Table of Contents
King, Steven R; Duggal, Nisha K; Ndongmo, Clement B et al. (2008) Pseudodiploid genome organization AIDS full-length human immunodeficiency virus type 1 DNA synthesis. J Virol 82:2376-84
Duggal, Nisha K; Goo, Leslie; King, Steven R et al. (2007) Effects of identity minimization on Moloney murine leukemia virus template recognition and frequent tertiary template-directed insertions during nonhomologous recombination. J Virol 81:12156-68
Takebe, Yutaka; Telesnitsky, Alice (2006) Evidence for the acquisition of multi-drug resistance in an HIV-1 clinical isolate via human sequence transduction. Virology 351:1-6
Onafuwa-Nuga, Adewunmi A; Telesnitsky, Alice; King, Steven R (2006) 7SL RNA, but not the 54-kd signal recognition particle protein, is an abundant component of both infectious HIV-1 and minimal virus-like particles. RNA 12:542-6
An, Wenfeng; Telesnitsky, Alice (2004) Human immunodeficiency virus type 1 transductive recombination can occur frequently and in proportion to polyadenylation signal readthrough. J Virol 78:3419-28