Study Of Interaction Of HIV Proteins With Host Cells Using A Fission Yeast Model And Potential Strategy Against HIV

During the infection process, human immunodeficiency virus (HTV) may change the situation of host cells to make the cellular environment suitable for infection. However, upon HIV-1 infection host cells react as various innate, cellular and humoral immune responses to counteract the viral invasion. HIV-1 overcomes these antiviral responses through many different ways. Infection of HIV-1 is a process of host-pathogen interaction. Studying the interaction of viral proteins with host cells is important for understanding HIV infection mechanism and searching new potential anti-HIV strategy. Considering the complexity of mammalian cells, we have used fission yeast (S. pombe) as a model system to study the interaction of viral proteins with host cells and try to find new anti-HIV strategy.Part I. Molecular Characterization of HIV-1 genome by using fission yeast (S.pombe ) as a model systemAs a fungus, S. pombe has many features mimicking mammalian cells. Most of proteins have same functions in S.pombe cells and mammalian cells. Comparing with the complication of mammalian cells, S.pombe provides a good model system for molecular genetic studies of eukaryotic genes. The structural and functional information of eukaryotic genes, especially those of mammalian origin, provided by the research in S. pombe give us important insight into their mechanisms in humans. The study with S. pombe as a model system on HIV now is mainly focused on the Vpr protein and LTR promoter. In this thesis, we have expressed all of the HIV-1 proteins in S. pombe to investigate the subcellular localization of each protein and their effects on the growth of S. pombe cells. Based on those data, we tried to develop new screen systems to screen anti-HIV-1 inhibitors. Major results are listed below:A. Subcellular localizations of all of HIV-1 proteins were observed in S.pombe cells, the results have shown that gag protein accumulated into high light dot in cytoplasm; p24, IN, Vif and Rev localized in the nuclear; P7, P6 and Vpu localized in cytoplasm; Vpr localized on the nuclear membrane; the other proteins localized both in cytoplasm and nuclear.B. All of HIV-1 proteins were expressed separately in S.pombe cells and the effect of each protein on the growth of S.pombe cells was observed. The results have shown that HIV-1 Rev and PR expression delayed the growth of S.pombe cells.C. The growth delay caused by HTV Rev in S.pombe was not due to the cell death or cell cycle arrest.D. Rev protein has the same effect on the cell growth in mammalian cells as in...