Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/48334
Title: Survey of cellular factors modulating the HIV-1 integration complex activity using a unique protein screening system in vitro
Authors: TAN BENG HUI
Keywords: HIV-1 integration, HIV-1 integrase, pre-integration complex, E3 ubiquitin ligase, RFPL3, cellular host factors
Issue Date: 30-Aug-2013
Source: TAN BENG HUI (2013-08-30). Survey of cellular factors modulating the HIV-1 integration complex activity using a unique protein screening system in vitro. ScholarBank@NUS Repository.
Abstract: HIV-1 integration is mediated by preintegration complex (PIC) consisting of viral DNA associated with viral and cellular proteins required for early phase of infection. A striking feature of PIC is its powerful integration activity in vitro: PICs freshly isolated from infected cells are able to insert viral DNA into exogenously added target DNA, indicating that PIC-based assay allows a reliable assessment of retroviral integration. In this study, we developed an in vitro assay platform monitoring PIC-mediated integration in microtiter plates. In combination with a library of human E3 ubiquitin ligases generated by wheat germ cell-free system, we found that RFPL3 stimulated integration activity of HIV-1 PIC in vitro. This enhancement of PIC activity by RFPL3 was likely to be attributed to its N-terminal RING domain. Additionally, immunoprecipitation analysis revealed that RFPL3 was associated with HIV-1 PICs in infected cells. The level of viral infectivity was also higher in RFPL3-expressing cells.
URI: http://scholarbank.nus.edu.sg/handle/10635/48334
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