Raghavendra, Nidhanapati K. and Shkriabai, Nikolozi and Graham, Robert L. J. and Hess, Sonja and Kvaratskhelia, Mamuka and Wu, Li (2010) Identification of host proteins associated with HIV-1 preintegration complexes isolated from infected CD4^+ cells. Retrovirology, 7 . Art. No. 66. ISSN 1742-4690 http://resolver.caltech.edu/CaltechAUTHORS:20101027-100244817
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An integrated HIV-1 genomic DNA leads to an infected cell becoming either an active or a latent virus-producing cell. Upon appropriate activation, a latently infected cell can result in production of progeny viruses that spread the infection to uninfected cells. The host proteins influence several steps of HIV-1 infection including formation of the preintegration complex (PIC), a key nucleoprotein intermediate essential for integration of reverse transcribed viral DNA into the chromosome. Much effort has gone into the identification of host proteins contributing to the assembly of functional PICs. Experimental approaches included the use of yeast two-hybrid system, co-immunoprecipitation, affinity tagged HIV-1 viral proteins and in vitro reconstitution of salt-stripped PIC activity. Several host proteins identified using these approaches have been shown to affect HIV-1 replication in cells and influence catalytic activities of recombinant IN in vitro. However, the comprehensive identification and characterization of host proteins associated with HIV-1 PICs of infected cells have been hindered in part by the technical limitation in acquiring sufficient amount of catalytically active PICs. To efficiently identify additional host factors associated with PICs in infected cells, we have developed the following novel approach. The catalytically active PICs from HIV-1-infected CD4^+ cells were isolated using biotinylated target DNA, and the proteins selectively co-purifying with PICs have been analyzed by mass spectrometry. This technology enabled us to reveal at least 19 host proteins that are associated with HIV-1 PICs, of which 18 proteins have not been described previously with respect to HIV-1 integration. Physiological functions of the identified proteins range from chromatin organization to protein transport. A detailed characterization of these host proteins could provide new insights into the mechanism of HIV-1 integration and uncover new antiviral targets to block HIV-1 integration.
|Additional Information:||© 2010 Raghavendra et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Received: 31 May 2010 Accepted: 11 August 2010. Published: 11 August 2010. The following reagents were obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH: H9 and H9/HTLVIIIB cells from Dr. Robert Gallo. We thank Dr. Kathleen Boris-Lawrie for generous gift of antibodies used in the immunoblotting. This work was supported in part by grants to LW (R01AI068493 and R21AI078762) and to MK (R01AI062520 and P01CA100730) from the NIH. Authors’ contributions: NKR conceived the study, designed and performed the biochemical experiments and drafted the manuscript. SH and MK designed, and NS and RLJG performed the mass spectrometry and helped in drafting the manuscript. LW coordinated the study, participated in the experimental design and the drafting of the manuscript. All authors read and approved the final manuscript.|
|Official Citation:||Raghavendra et al.: Identification of host proteins associated with HIV-1 preintegration complexes isolated from infected CD4+ cells. Retrovirology 2010 7:66.|
|Usage Policy:||This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.|
|Deposited By:||Tony Diaz|
|Deposited On:||23 Nov 2010 22:37|
|Last Modified:||26 Dec 2012 12:34|
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