以裂殖酵母为模型研究HIV蛋白与宿主细胞的相互作用并探索抗HIV-1的新策略
发布时间：2018-08-23 19:31
【摘要】：人类免疫缺陷病毒(Human Immunodeficiency Virus，HIV)在感染宿主细胞的过程中通过多种方式改变细胞状态使其适合于病毒复制，而宿主细胞则利用特异性或非特异性的反应来抵抗HIV的感染，HIV必须克服这些抗病毒反应以保证其正常复制，因此可以说HIV感染的过程就是一个病毒蛋白与宿主相互作用的过程。研究HIV病毒蛋白与宿主的相互作用是阐明HIV病毒感染机制、探讨新的抗病毒方法的基础。鉴于人细胞的复杂性，我们利用S．pombe细胞为模型研究HIV病毒蛋白与宿主细胞蛋白的相互作用，并以此为基础探讨新的抗HIV-1策略。 第一部分 以S．pombe为模型对HIV-1全基因组的研究 S．pombe是与哺乳动物细胞非常相似的一种真菌，各种真核细胞蛋白在S．pombe细胞和哺乳动物细胞中的功能基本相似。因此，S．pombe细胞为真核细胞蛋白结构、功能、基因表达的研究提供了很好的模型。在S．pombe细胞中得到的实验数据能够为真核细胞中的机制研究提供很好的启示，从而指导复杂的哺乳动物细胞中的研究。S．pombe细胞目前已经被应用于HIV病毒蛋白功能的研究，主要是集中于HIV-1 Vpr和LTR启动子的研究。我们将HIV-1所有共17个蛋白在S．pombe细胞中进行了表达，观察了17种蛋白在S．pombe细胞中的定位及各蛋白表达对S．pombe细胞生长的影响，并将结果与哺乳动物细胞中的数据进行了比较，在此基础上探讨了建立筛选抗HIV-1蛋白功能抑制剂系统的可行性，主要结果如下： A．在S．pombe细胞中观察了GFP融合的HIV-1所有17种蛋白的细胞内的定位，，发现gag蛋白在细胞内聚集呈点状分布，P24、IN、Vif和Rev定位于细胞核中，P7、P6、Vpu、位于细胞浆中，Vpr位于细胞膜上，其它蛋白在细胞核和细胞浆中均有分布。 B．在S．pombe细胞中表达了HIV-1的所有蛋白，发现Rev蛋白和PR蛋白表达能够导致S．pombe细胞的生长缓慢。
[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.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R373
本文编号：2199692
[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.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R373
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