The objective of this proposal is to identify the function of HIV-1 Vif and determine the role of Vif in the virus life cycle. The long-term objective of these studies is to identify new molecular mechanisms in the HIV-1 virus life cycle which could potentially serve as targets for antiviral therapy. Vif is potentially an excellent target for antiviral therapy since its function is essential for HIV-1 replication in peripheral blood T lymphocytes and monocyte/macrophages. There are currently no assay to screen drugs which inhibit Vif, since its mechanism of action and active sites are unknown. Vif-defective viruses enter cells normally but are defective in their ability to complete proviral DNA synthesis. Recently, we have shown that Vif-defective virions show an abnormality in the structure of the virion core. More significantly, we have examined the effect of Vif on the endogenous reverse transcriptase reaction in vitro and have identified a major defect in the ability of vif-defective virions to synthesize viral DNA using endogenous viral RNA as a template. These results are significant because they indicate that a component of the virion core, such as the viral RNA genome or one of the gag or gag-pol gene products, is likely to be an important target for Vif function. The endogenous reverse transcriptase reaction and other molecular approaches will be used to identify the mechanism of the effect of Vif on proviral DNA synthesis. These in vitro studies will be complemented by studies in cell lines and primary cells to determine biological significance during early events in vivo. Studies on the role of Vif in virus assembly and maturation will include examination of the effect of Vif on the viral RNA genome and viral RNA-nucleocapsid interactions and the possibility of a direct interaction between Vif and Gag during virus production. Vif mutants in an infectious molecular clone will be used to determine the biological significance of Vif phenotypes during virus replication and to integrate data from the different sections of this proposal. Additional insights into the biochemistry and cell biology of Vif resulting from other ongoing studies in the laboratory will be used to assess new phenotypes of Vif as the project progresses. These studies may lead to the identification of mechanisms relevant ot Vif function which could potentially serve as targets for therapeutic intervention. The broad range of studies on several critical steps in the retrovirus life cycle, including reverse transcription, formation of the DNA provirus, and the organization of the viral RNA genome in the virion, might also provide insights which may be relevant for the development of retroviral vectors.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI036186-04
Application #
2672342
Study Section
AIDS and Related Research Study Section 3 (ARRC)
Project Start
1995-04-01
Project End
1999-07-31
Budget Start
1998-04-01
Budget End
1999-07-31
Support Year
4
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Dana-Farber Cancer Institute
Department
Type
DUNS #
149617367
City
Boston
State
MA
Country
United States
Zip Code
02215
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Mehle, Andrew; Goncalves, Joao; Santa-Marta, Mariana et al. (2004) Phosphorylation of a novel SOCS-box regulates assembly of the HIV-1 Vif-Cul5 complex that promotes APOBEC3G degradation. Genes Dev 18:2861-6
Mehle, Andrew; Strack, Bettina; Ancuta, Petronela et al. (2004) Vif overcomes the innate antiviral activity of APOBEC3G by promoting its degradation in the ubiquitin-proteasome pathway. J Biol Chem 279:7792-8
Aires da Silva, Frederico; Santa-Marta, Mariana; Freitas-Vieira, Acilino et al. (2004) Camelized rabbit-derived VH single-domain intrabodies against Vif strongly neutralize HIV-1 infectivity. J Mol Biol 340:525-42
Goncalves, Joao; Silva, Frederico; Freitas-Vieira, Acilino et al. (2002) Functional neutralization of HIV-1 Vif protein by intracellular immunization inhibits reverse transcription and viral replication. J Biol Chem 277:32036-45
Ohagen, A; Gabuzda, D (2000) Role of Vif in stability of the human immunodeficiency virus type 1 core. J Virol 74:11055-66
Yang, X; Gabuzda, D (1999) Regulation of human immunodeficiency virus type 1 infectivity by the ERK mitogen-activated protein kinase signaling pathway. J Virol 73:3460-6
Yang, X; Chen, Y; Gabuzda, D (1999) ERK MAP kinase links cytokine signals to activation of latent HIV-1 infection by stimulating a cooperative interaction of AP-1 and NF-kappaB. J Biol Chem 274:27981-8
Yang, X; Gabuzda, D (1998) Mitogen-activated protein kinase phosphorylates and regulates the HIV-1 Vif protein. J Biol Chem 273:29879-87
Hoglund, S; Ohagen, A; Goncalves, J et al. (1997) Ultrastructure of HIV-1 genomic RNA. Virology 233:271-9

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