F11-mediated inhibition of RhoA signalling enhances the spread of vaccinia virus in vitro and in vivo in an intranasal mouse model of infection.
The cortical actin cytoskeleton beneath the plasma membrane represents a physical barrier that vaccinia virus has to overcome during its exit from an infected cell. Previous observations using overexpression and pharmacological approaches suggest that vaccinia enhances its release by modulating the cortical actin cytoskeleton by inhibiting RhoA signalling using the viral protein F11. We have now examined the role of F11 and its ability to interact with RhoA to inhibit its downstream signalling in the spread of vaccinia infection both in vitro and in vivo. Live cell imaging over 48 hours reveals that loss of F11 or its ability to bind RhoA dramatically reduces the rate of cell-to-cell spread of the virus in a cell monolayer. Cells infected with the DeltaF11L virus also maintained their cell-to-cell contacts, and did not undergo virus-induced motility as observed during wild-type infections. The DeltaF11L virus is also attenuated in intranasal mouse models of infection, as it is impaired in its ability to spread from the initial sites of infection to the lungs and spleen. Loss of the ability of F11 to bind RhoA also reduces viral spread in vivo. Our results clearly establish that viral-mediated inhibition of RhoA signalling can enhance the spread of infection not only in cell monolayers, but also in vivo.
Cordeiro
João V
JV
Cell Motility Laboratory, London Research Institute, London, United Kingdom.
Guerra
Susana
S
Arakawa
Yoshiki
Y
Dodding
Mark P
MP
Esteban
Mariano
M
Way
Michael
M
D000199:Actins / Q000378:metabolism; D000281:Administration, Intranasal; D000818:Animals; D002465:Cell Movement; D004195:Disease Models, Animal; D006160:Guanosine Triphosphate / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D011485:Protein Binding; D015398:Signal Transduction; D022502:Stress Fibers / Q000378:metabolism / Q000821:virology; D014615:Vaccinia / Q000821:virology*; D014616:Vaccinia virus / Q000502:physiology*; D014764:Viral Proteins / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
D000199:Actins; C533030:F11L protein, vaccinia virus; D014764:Viral Proteins; D006160:Guanosine Triphosphate; D020742:rhoA GTP-Binding Protein