Previous studies have shown that HIV-1 preferentially buds from cholesterol-rich microdomains called lipid rafts, and that cholesterol is required for HIV infectivity and structural integrity. Moreover, the presence of cholesterol is also required both for HIV entry into susceptible cells as well as for release of infectious virions. The viral requirement of cholesterol for HIV infectivity led us to study the relationship between HIV-1 budding and assembly and cholesterol trafficking. Niemann Pick Type C (NPC2) protein plays a vital role in the intracellular trafficking of cholesterol. Given the viral requirement of cholesterol for productivity and given the role of NPC2 in cholesterol trafficking, we hypothesize that NPC2 activity is required in the late stages of HIV-1 biogenesis. HIV-1 induced lipodystrophy and the consequent cardiovascular syndromes, as well as the critical role of cholesterol in viral pathogenesis, make this study highly relevant to NIH's mission to eliminate HIV/AIDS health disparities and to promote public health. The findings from this project may also offer insight into other cholesterol- associated disparities. To this end, the following specific aims have been developed: 1) To test whether or not NPC2 plays a role in HIV-1 replication. 2) To examine the mechanism by which cholesterol trafficking effects viral production using NPC disease as a model. The primary scientific techniques that will be utilized to address these aims are Western analysis, PCR, gene silencing, ELISA, flow cytometry, and microscopy.
| Coleman, Ebony M; Walker, Tiffany N; Hildreth, James E K (2012) Loss of Niemann Pick type C proteins 1 and 2 greatly enhances HIV infectivity and is associated with accumulation of HIV Gag and cholesterol in late endosomes/lysosomes. Virol J 9:31 |
