人泡沫病毒LTR与细胞锌指蛋白ZNF268相互作用机制及功能研究

人泡沫病毒（HFV）属逆转录病毒科，其独特的感染机制和不明显的致病作用使得HFV作为安全的基因转移载体而日益受到人们的重视。LTR是HFV的启动子，在病毒感染和复制中具有重要功能，但是其调控机制目前尚不十分清楚。我们前期工作通过EMSA实验筛选出HFV LTR的结合蛋白- - ZNF268蛋白，且luciferase分析方法初步证实了LTR的调控功能受ZNF268抑制；本项目将进一步结合序列突变分析确证它们的相互作用及结合位点，通过RNA干扰、免疫共沉淀、荧光共振能量转移和Western blotting等技术确定相互作用的信号转导途径与机制，深入探讨LTR与ZNF268相互作用对病毒感染、转录、复制、病毒滴度等功能的影响。本项目的完成将为阐明LTR新的调控功能，以及揭示HFV感染分子机制和载体安全性等都具有重要的意义，也将为探讨其它逆转录病毒的感染分子机制与抗病毒治疗提供有益的线索。

Human foamy virus (HFV) belongs to retroviridae. It is of great interest that HFV can be a promising safe vector system for gene transfer with its nonpathogenic nature and the unique features of infection mechanism. The long terminal repeat (LTR) of a HFV promoter, which directs the expression of the structural proteins and the regulatory proteins, plays an important role in viral infection and replication. However, it is largely unknown about the regulatory mechanism of LTR in viral infection and replication. Previously, we identified the ZNF268 protein that could bind HFV LTR by EMSA experiment. Moreover, it was found that the transcriptional function of LTR could be repressed by ZNF268 through the method of the luciferase analysis. In this study, their interaction and binding site between LTR and ZNF268 will be further confirmed by the method of sequence mutational analysis. Subsequently, the pathway and mechanism of the interaction between LTR and ZNF268 will be determined by the methods of RNA interference, Co-IP、Fluorescence resonance energy transfer and Western blotting. Furthermore, we will investigate how the interaction of LTR and ZNF268 affects viral infection, transcription, replication and Virus titer. The implement of this project will elucidate the new regulatory function of LTR and reveal HFV infecting molecular mechanism and vector safety. It also will provide the helpful clue for other retroviruses and anti-viral therapy.