细胞内源蛋白TTRAP与HIV-1整合酶的相互作用及其对慢病毒整合的影响
发布时间：2019-05-23 11:58
【摘要】：TTRAP是一个最近发现的与NF-kB通路相关的PML核体蛋白。在这项研究中,通过酵母双杂交分析,TTRAP被鉴定为一个HIV-1整合酶相互作用蛋白。这样的相互作用被进一步通过免疫共沉淀,GST pull down和细胞内共定位实验加以证明。并且,截断体分析表明,TTRAP的N端的180个氨基酸残基直接与HIV-1整合酶结合。在稳定敲除TTRAP的细胞系中,病毒载体的整合效率相比整合non-silencing载体的对照细胞系大大下降;而另一方面,感染病毒载体前瞬时转染pcDNA3.1-TTRAP质粒过表达TTRAP增加了整合事件发生的百分率。综上,本实验报道了细胞内蛋白TTRAP具有与HIV-1整合酶结合的能力,并且促进慢病毒载体的整合。这些发现揭示了TTRAP的一个新功能,并且拓展了我们关于细胞应答HIV-1感染的知识。TTRAP和HIV-1整合酶存在的相互作用可能帮助我们设计新的抗病毒药物和提高慢病毒介导的基因转移的效率。
[Abstract]:TTRAP is a recently discovered PML nucleosome protein related to NF-kB pathway. In this study, TTRAP was identified as a HIV-1 integrase interacting protein by yeast two-hybrid analysis. This interaction was further proved by immunoprecipitation, GST pull down and intracellular co-localization. Moreover, the analysis of truncated body showed that the N-terminal amino acid residues of TTRAP were directly bound to HIV-1 integrase. In the cell line stably knockout TTRAP, the integration efficiency of virus vector was much lower than that of the control cell line integrated with non-silencing vector. On the other hand, transient transfer of TTRAP into pcDNA3.1-TTRAP plasmid before infection of virus vector increased the percentage of integration events. In summary, this study reported that intracellular protein TTRAP has the ability to bind to HIV-1 integrase and promote the integration of lentivirus vector. These findings reveal a new feature of TTRAP. The interaction between TTRAP and HIV-1 integrase may help us design new antiviral drugs and improve the efficiency of lentivirus mediated gene transfer.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R341
本文编号：2483876
[Abstract]:TTRAP is a recently discovered PML nucleosome protein related to NF-kB pathway. In this study, TTRAP was identified as a HIV-1 integrase interacting protein by yeast two-hybrid analysis. This interaction was further proved by immunoprecipitation, GST pull down and intracellular co-localization. Moreover, the analysis of truncated body showed that the N-terminal amino acid residues of TTRAP were directly bound to HIV-1 integrase. In the cell line stably knockout TTRAP, the integration efficiency of virus vector was much lower than that of the control cell line integrated with non-silencing vector. On the other hand, transient transfer of TTRAP into pcDNA3.1-TTRAP plasmid before infection of virus vector increased the percentage of integration events. In summary, this study reported that intracellular protein TTRAP has the ability to bind to HIV-1 integrase and promote the integration of lentivirus vector. These findings reveal a new feature of TTRAP. The interaction between TTRAP and HIV-1 integrase may help us design new antiviral drugs and improve the efficiency of lentivirus mediated gene transfer.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R341
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