Die Rolle von UL128-131-abhängigen Endozytosemechanismen für den Endothelzelltropismus des menschlichen Cytomegalovirus (HCMV)

该项目基于以下假设：人巨细胞病毒（HCMV）的被膜或衣壳蛋白与动力蛋白/动力肌动蛋白复合物的细胞蛋白的相互作用对于嗜内皮HCMV/E株和非嗜内皮HCMV/F株之间的细胞嗜性差异至关重要。我们将（1）通过构建具有相应HCMV/E蛋白的假型化HCMV/F病毒来分析各种病毒结构蛋白对EC嗜性的贡献，（2）将联合收割机单病毒体免疫荧光染色与细胞骨架组分的差异去除相结合，以剖析肌动蛋白和微管在病毒穿透、向MTOC运输和递送到细胞核期间的作用，和（3）通过将差异沉淀与2-D凝胶电泳和蛋白质鉴定技术相结合来分析病毒和细胞结构蛋白之间的结合和磷酸化/去磷酸化事件。总之，我们期望深入了解HCMV EC嗜性菌株间差异的分子机制，这对开发新的抗病毒策略和开发EC靶向疫苗具有潜在意义。

This project is based on the hypothesis that interactions of tegument or capsid proteins of human cytomegalovirus (HCMV) with cellular proteins of the dynein/dynactin complex are critical for cell tropism differences between endotheliotropic HCMV/E strains and nonendotheliotropic HCMV/F strains. We will (1) analyze the contribution of various viral structural proteins to EC tropism by the construction of HCMV/F viruses pseudotyped with the respective HCMV/E proteins, (2) combine single virion immunofluorescence staining with differential removal of cytoskeleton components to dissect the role of actin and microtubules during viral penetration, transport towards the MTOC and delivery to the nucleus, and (3) analyze events of binding and phosphorylation/dephosphorylation between viral and cellular structural proteins by a combination of differential precipitation with 2-D gel electrophoresis and protein identification techniques. In conclusion, we expect insight into the molecular mechanisms underlying interstrain differences in the EC tropism of HCMV, with potential implications for the development of new antiviral strategies and the development of EC-targeted vaccines.