The Vif protein is highly conserved among lentiviruses, and is crucial for HIV-1 replication in primary T lymphocytes and macrophages, the cells infected by the virus in vivo. Our work has demonstrated that Vif is necessary at the time of viral particle formation, to make HIV-1 competent for proviral DNA synthesis. Furthermore, our recent analyses indicate that Vif stabilizes the HIV-1 reverse transcription complex in vitro. Progress in the Vif field has been slowed down by the technical difficulty of obtaining high-titer stocks of functionally relevant virus. We will use two parallel approaches to solve this problem. This will facilitate the further exploration of VIF function and mechanisms of action, using a combination of functional and biochemical analyses. Finally, the search for viral and cellular proteins interacting with Vif will complement these studies.
our specific aims are thus as follows: 1) Creation of regulated vif expression systems, utilizing a tetracycline- repressible and an ecdysone-inducible system. 2) Generation of conditional vif mutants, by site-directed and saturation mutagenesis. 3) Continuing studies of Vif function and mechanism of action, by further analyzing the Vif-mediated enhancement of reverse transcription, studying the processing and post-translational modification of virion-associated proteins, and examining the early steps of infection through new protocols developed in our laboratory. 4) Search for viral and/or cellular proteins that associate with Vif, by co-immunoprecipitation and using the yeast two-hybrid system.
| Camaur, D; Trono, D (1996) Characterization of human immunodeficiency virus type 1 Vif particle incorporation. J Virol 70:6106-11 |
| von Schwedler, U; Kornbluth, R S; Trono, D (1994) The nuclear localization signal of the matrix protein of human immunodeficiency virus type 1 allows the establishment of infection in macrophages and quiescent T lymphocytes. Proc Natl Acad Sci U S A 91:6992-6 |
| von Schwedler, U; Song, J; Aiken, C et al. (1993) Vif is crucial for human immunodeficiency virus type 1 proviral DNA synthesis in infected cells. J Virol 67:4945-55 |
