While the essential nucleic acid chemistry of the integration reaction is reasonably understood, how integration of the viral pre-integration complex actually happens in the complex environment of the cell nucleus remains mysterious in many respects. In particular, the integration field is intensely interested in how cellular molecules participate either positively or negatively in the reaction between the pre-integration complex and chromatin, and also in how they determine where and when the reaction occurs in the host cell genome. The transcriptional coactivator LEDGF/p75 has now been securely implicated as an integration cofactor specific for lentiviruses. A basic molecular tethering model has been put forth in which LEDGF/p75 tethers IN to chromatin, key participating protein domains have been identified, and precise structural information exists for the LEDGF/p75 domain that interacts with IN. Hypothesized roles of LEDGF/p75 include acting as a cofactor for catalysis steps, protecting IN and the pre- integration complex in which it resides from degradation, and guiding or tethering the pre-integration complex to chromatin. The latter mechanism could be crucial for the virus to engage chromatin and integrate per se, and it also may determine the distinctive genomic distributions now evident for HIV integration sites. Understanding the importance of LEDGF/p75 to HIV biology may therefore have long-term implications for understanding latency and may provide a new therapeutic target. It could even pave the way for targeting of lentiviral vectors to reduce insertional mutagenesis risks. In this project, we will use effective RNAi knockdown methods, genetic knockout cells, virological analyses, domain mapping and heterologous tethering techniques, genome- wide integration site analyses, and cell biological and proteomic methods to both further understand and exploit the viral biology of LEDGF/p75.

Public Health Relevance

This project will elucidate the significance of the transcriptional cofactor LEDGF/p75 in the HIV-1 life cycle, using virological, biochemical, genomic and chimeric protein approaches.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
3R01AI077344-04S1
Application #
8433105
Study Section
Special Emphasis Panel (ZRG1-AARR-D (05))
Program Officer
Salzwedel, Karl D
Project Start
2008-05-01
Project End
2013-04-30
Budget Start
2012-03-01
Budget End
2012-04-30
Support Year
4
Fiscal Year
2012
Total Cost
$7,846
Indirect Cost
$2,871
Name
Mayo Clinic, Rochester
Department
Type
DUNS #
006471700
City
Rochester
State
MN
Country
United States
Zip Code
55905
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Thierry, Sylvain; Munir, Soundasse; Thierry, Eloïse et al. (2015) Integrase inhibitor reversal dynamics indicate unintegrated HIV-1 dna initiate de novo integration. Retrovirology 12:24
Painter, Meghan M; Morrison, James H; Zoecklein, Laurie J et al. (2015) Antiviral Protection via RdRP-Mediated Stable Activation of Innate Immunity. PLoS Pathog 11:e1005311
Singh, Parmit Kumar; Plumb, Matthew R; Ferris, Andrea L et al. (2015) LEDGF/p75 interacts with mRNA splicing factors and targets HIV-1 integration to highly spliced genes. Genes Dev 29:2287-97
Slaughter, Alison; Jurado, Kellie A; Deng, Nanjie et al. (2014) The mechanism of H171T resistance reveals the importance of Nδ-protonated His171 for the binding of allosteric inhibitor BI-D to HIV-1 integrase. Retrovirology 11:100
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Sharma, Amit; Slaughter, Alison; Jena, Nivedita et al. (2014) A new class of multimerization selective inhibitors of HIV-1 integrase. PLoS Pathog 10:e1004171
Morrison, James H; Guevara, Rebekah B; Marcano, Adriana C et al. (2014) Feline immunodeficiency virus envelope glycoproteins antagonize tetherin through a distinctive mechanism that requires virion incorporation. J Virol 88:3255-72
Xu, Xin; Powell, David W; Lambring, Courtney J et al. (2014) Human MCS5A1 candidate breast cancer susceptibility gene FBXO10 is induced by cellular stress and correlated with lens epithelium-derived growth factor (LEDGF). Mol Carcinog 53:300-13
Meehan, Anne M; Saenz, Dyana T; Guevera, Rebekah et al. (2014) A cyclophilin homology domain-independent role for Nup358 in HIV-1 infection. PLoS Pathog 10:e1003969

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