vesicular stomatitis virus enters cells through vesicles incompletely coated with clathrin that depend upon actin for internalization水泡性口炎病毒进入细胞通过囊泡不完全覆盖的网格蛋白依赖肌动蛋白内化.pdfVIP

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vesicular stomatitis virus enters cells through vesicles incompletely coated with clathrin that depend upon actin for internalization水泡性口炎病毒进入细胞通过囊泡不完全覆盖的网格蛋白依赖肌动蛋白内化.pdf

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vesicular stomatitis virus enters cells through vesicles incompletely coated with clathrin that depend upon actin for internalization水泡性口炎病毒进入细胞通过囊泡不完全覆盖的网格蛋白依赖肌动蛋白内化

Vesicular Stomatitis Virus Enters Cells through Vesicles Incompletely Coated with Clathrin That Depend upon Actin for Internalization David K. Cureton1,2, Ramiro H. Massol3,4,5, Saveez Saffarian4,5, Tomas L. Kirchhausen4,5*, Sean P. J. Whelan1,2* 1 Departments of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts, United States of America, 2 Program in Virology, Harvard Medical School, Boston, Massachusetts, United States of America, 3 Children’s Hospital, Boston, Massachusetts, United States of America, 4 Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, United States of America, 5 Immune Disease Institute, Harvard Medical School, Boston, Massachusetts, United States of America Abstract Many viruses that enter cells by clathrin-dependent endocytosis are significantly larger than the dimensions of a typical clathrin-coated vesicle. The mechanisms by which viruses co-opt the clathrin machinery for efficient internalization remain uncertain. Here we examined how clathrin-coated vesicles accommodate vesicular stomatitis virus (VSV) during its entry into cells. Using high-resolution imaging of the internalization of single viral particles into cells expressing fluorescent clathrin and adaptor molecules, we show that VSV enters cells through partially clathrin-coated vesicles. We found that on average, virus-containing vesicles contain more clathrin and clathrin adaptor molecules than conventional vesicles, but this increase is insufficient to permit full coating of the vesicle. We further show that virus-containing vesicles depend upon the actin machinery for their internalization. Specifically, we found that components of the actin machinery are recruited to virus-containing vesicles, and chemical inhibition of actin polymerization trapped viral particles in vesicles at the plasma membrane. By analysis of multiple independe