Role of clathrin-mediated endocytosis during vesicular stomatitis virus entry into host cells

Abstract

Vesicular stomatitis virus (VSV) is well established to enter cells by pH-dependent endocytosis, but mechanistic aspects of its internalization have remained unclear. Here, we examined the functional role of clathrin in VSV entry by expression of a dominant-negative mutant of Eps15 (GFP-Eps15Delta95/295), a protein essential for clathrin-mediated endocytosis. Whereas expression of GFP alone had no effect on VSV infection, expression of GFP-Eps15Delta95/295 severely limited infection. As independent ways to examine clathrin function, we also examined cells that had been treated with chlorpromazine and utilized small interfering RNA (siRNA) techniques. Inhibition of clathrin-mediated endocytosis by chlorpromazine treatment, as well as clathrin knock-down using siRNA duplexes directed against the clathrin heavy chain, also prevented VSV infection. In combination with previous morphological approaches, these experiments establish clathrin as an essential component needed for endocytosis of VSV.

Involvement of a Rac1-Dependent Macropinocytosis Pathway in Plasmid DNA Delivery by Electrotransfection

“…8,83,85,86 Vesicles in clathrin-and caveolin-mediated endocytosis are small in size (<100 nm), which can significantly limit their transport capacities for pDNA with a similar size. In contrast, macropinocytosis, which is known to be responsible for cellular uptake of viruses, 87,88 nanoparticles, 89 and naked pDNA, 90 can generate vesicles with sizes of up to 5 mm that could transport a significantly larger amount of pDNA per vesicle. In previous and current studies, it was observed that pDNA could form aggregates in solution with certain sizes or complexes with the plasma membrane.…”

Visualization of targeted transduction by engineered lentiviral vectors

“…42 It has been shown that the fusion of VSV occurs before the late endosome stage. 27 Our data suggest that it takes 70-80 min more for the engineered lentivirus to reach a half-maximum of fusion-involved penetration than it does for VSVG-pseudotyped virus.…”

RNAi screening reveals requirement for host cell secretory pathway in infection by diverse families of negative-strand RNA viruses

“…During infection, VSV binds to susceptible cells, although the receptor(s) mediating virus entry remains unidentified (3). It enters cells by clathrin-mediated endocytosis, requiring endocytic adaptor protein AP-2, actin, and dynamin (4)(5)(6). Once in the cytoplasm, low pH-dependent fusion of viral envelope with the endosomal membrane leads to the release of NC in the cytoplasm for transcription and replication to occur.…”