【摘要】：人類免疫缺陷病毒(Human Immunodeficiency Virus，HIV)在感染宿主細胞的過程中通過多種方式改變細胞狀態(tài)使其適合于病毒復制，而宿主細胞則利用特異性或非特異性的反應來抵抗HIV的感染，HIV必須克服這些抗病毒反應以保證其正常復制，因此可以說HIV感染的過程就是一個病毒蛋白與宿主相互作用的過程。研究HIV病毒蛋白與宿主的相互作用是闡明HIV病毒感染機制、探討新的抗病毒方法的基礎(chǔ)。鑒于人細胞的復雜性，我們利用S．pombe細胞為模型研究HIV病毒蛋白與宿主細胞蛋白的相互作用，并以此為基礎(chǔ)探討新的抗HIV-1策略。 第一部分 以S．pombe為模型對HIV-1全基因組的研究 S．pombe是與哺乳動物細胞非常相似的一種真菌，各種真核細胞蛋白在S．pombe細胞和哺乳動物細胞中的功能基本相似。因此，S．pombe細胞為真核細胞蛋白結(jié)構(gòu)、功能、基因表達的研究提供了很好的模型。在S．pombe細胞中得到的實驗數(shù)據(jù)能夠為真核細胞中的機制研究提供很好的啟示，從而指導復雜的哺乳動物細胞中的研究。S．pombe細胞目前已經(jīng)被應用于HIV病毒蛋白功能的研究，主要是集中于HIV-1 Vpr和LTR啟動子的研究。我們將HIV-1所有共17個蛋白在S．pombe細胞中進行了表達，觀察了17種蛋白在S．pombe細胞中的定位及各蛋白表達對S．pombe細胞生長的影響，并將結(jié)果與哺乳動物細胞中的數(shù)據(jù)進行了比較，在此基礎(chǔ)上探討了建立篩選抗HIV-1蛋白功能抑制劑系統(tǒng)的可行性，主要結(jié)果如下： A．在S．pombe細胞中觀察了GFP融合的HIV-1所有17種蛋白的細胞內(nèi)的定位，，發(fā)現(xiàn)gag蛋白在細胞內(nèi)聚集呈點狀分布，P24、IN、Vif和Rev定位于細胞核中，P7、P6、Vpu、位于細胞漿中，Vpr位于細胞膜上，其它蛋白在細胞核和細胞漿中均有分布。 B．在S．pombe細胞中表達了HIV-1的所有蛋白，發(fā)現(xiàn)Rev蛋白和PR蛋白表達能夠?qū)е耂．pombe細胞的生長緩慢。
[Abstract]:Human immunodeficiency virus (Human Immunodeficiency Virus,HIV) changes the state of a host cell in a variety of ways to make it suitable for viral replication. The host cells use specific or non-specific reactions to resist HIV infection and must overcome these antiviral responses to ensure their normal replication. Therefore, it can be said that the process of HIV infection is a process of interaction between virus protein and host. The study of the interaction between HIV virus protein and host is the basis for elucidating the mechanism of HIV virus infection and exploring new antiviral methods. In view of the complexity of human cells, we use S.pombe cells as a model to study the interaction between HIV virus proteins and host cell proteins, and explore a new anti-HIV-1 strategy. The first part is a study of the whole genome of HIV-1 using S.pombe as a model. S.pombe is a fungus that is very similar to mammalian cells. The functions of various eukaryotic proteins are similar in S.pombe cells and mammalian cells. Therefore, S. pombe cells provide a good model for the study of protein structure, function and gene expression in eukaryotic cells. The experimental data obtained in S.pombe cells can provide good inspiration for the study of the mechanism in eukaryotic cells, and thus guide the study of complex mammalian cells. S. pombe cells have been applied to the study of the function of HIV virus protein. It mainly focuses on the study of HIV-1 Vpr and LTR promoter. All 17 proteins of HIV-1 were expressed in S.pombe cells. The localization of 17 proteins in S.pombe cells and the effect of expression of 17 proteins on the growth of S.pombe cells were observed. The results were compared with the data in mammalian cells, and the feasibility of screening anti HIV-1 protein functional inhibitor system was discussed. The main results were as follows: A. the intracellular localization of all 17 proteins of HIV-1 fused with GFP was observed in S.pombe cells. It was found that the gag protein was located in the nucleus of P7, P6, and in the cytoplasm of the cell membrane. Other proteins are distributed in nucleus and cytoplasm. B. all proteins of HIV-1 are expressed in S.pombe cells. It was found that the expression of Rev protein and PR protein resulted in slow growth of S.pombe cells.
【學位授予單位】：中國人民解放軍軍事醫(yī)學科學院
【學位級別】：博士
【學位授予年份】：2006
【分類號】：R373

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