Host cell factors and functions involved in vesicular stomatitis virus entry.
Vesicular stomatitis virus (VSV) is an animal virus that based on electron microscopy and its dependence on acidic cellular compartments for infection is thought to enter its host cells in a clathrin-dependent manner. The exact cellular mechanism, however, is largely unknown. In this study, we characterized the entry kinetics of VSV and elucidated viral requirements for host cell factors during infection in HeLa cells. We found that endocytosis of VSV was a fast process with a half time of 2.5 to 3 min and that acid activation occurred within 1 to 2 min after internalization in early endosomes. The majority of viral particles were endocytosed in a clathrin-based, dynamin-2-dependent manner. Although associated with some of the surface-bound viruses, the classical adaptor protein complex AP-2 was not required for infection. Time-lapse microscopy revealed that the virus either entered preformed clathrin-coated pits or induced de novo formation of pits. Dynamin-2 was recruited to plasma membrane-confined virus particles. Thus, VSV can induce productive internalization by exploiting a specific combination of the clathrin-associated proteins and cellular functions.
Johannsdottir
Hrefna Kristin
HK
Institute of Biochemistry, ETH Zurich, Schafmattstrasse 18, CH-8093 Zurich, Switzerland.
Mancini
Roberta
R
Kartenbeck
Jurgen
J
Amato
Lea
L
Helenius
Ari
A
D033962:Adaptor Protein Complex 2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D002966:Clathrin / Q000037:antagonists & inhibitors / Q000378:metabolism; D034285:Dynamin II / Q000037:antagonists & inhibitors / Q000378:metabolism; D004705:Endocytosis; D011992:Endosomes / Q000821:virology; D004847:Epithelial Cells / Q000648:ultrastructure / Q000821:virology; D006367:HeLa Cells; D006801:Humans; D046529:Microscopy, Electron, Transmission; D018715:Microscopy, Video; D013997:Time Factors; D018116:Vesiculovirus / Q000502:physiology*; D053586:Virus Internalization
D033962:Adaptor Protein Complex 2; D002966:Clathrin; D034285:Dynamin II