Alphavirus Genome Delivery Occurs Directly at the Plasma Membrane in a Time- and Temperature-Dependent Process
0303 health sciences
Time Factors
Staining and Labeling
Cell Membrane
Temperature
Virus Internalization
3. Good health
03 medical and health sciences
Cricetinae
Animals
RNA, Viral
Sindbis Virus
Microscopy, Immunoelectron
Cells, Cultured
DOI:
10.1128/jvi.03412-12
Publication Date:
2013-02-07T02:58:00Z
AUTHORS (5)
ABSTRACT
ABSTRACT
It is widely held that arboviruses such as the alphavirus Sindbis virus gain entry into cells by a process of receptor-mediated endocytosis followed by membrane fusion in the acid environment of the endosome. We have used an approach of direct observation of Sindbis virus entry into cells by electron microscopy and immunolabeling of virus proteins with antibodies conjugated to gold beads. We found that upon attaching to the cell surface, intact RNA-containing viruses became empty shells that could be identified only by antibody labeling. We found that the rate at which full particles were converted to empty particles increased with time and temperature. We found that this entry event takes place at temperatures that inhibit both endosome formation and membrane fusion. We conclude that entry of alphaviruses is by direct penetration of cell plasma membranes through a pore structure formed by virus and, possibly, host proteins.
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