Retroviral assembly and budding occur through an actin-driven mechanism.
The assembly and budding of a new virus is a fundamental step in retroviral replication. Yet, despite substantial progress in the structural and biochemical characterization of retroviral budding, the underlying physical mechanism remains poorly understood, particularly with respect to the mechanism by which the virus overcomes the energy barrier associated with the formation of high membrane curvature during viral budding. Using atomic force, fluorescence, and transmission electron microscopy, we find that both human immunodeficiency virus and Moloney murine leukemia virus remodel the actin cytoskeleton of their host. These actin-filamentous structures assemble simultaneously with or immediately after the beginning of budding, and disappear as soon as the nascent virus is released from the cell membrane. Analysis of sections of cryopreserved virus-infected cells by transmission electron microscopy reveals similar actin filament structures emerging from every nascent virus. Substitution of the nucleocapsid domain implicated in actin binding by a leucine-zipper domain results in the budding of virus-like particles without remodeling of the cell's cytoskeleton. Notably, viruses carrying the modified nucleocapsid domains bud more slowly by an order of magnitude compared to the wild-type. The results of this study show that retroviruses utilize the cell cytoskeleton to expedite their assembly and budding.
Gladnikoff
Micha
M
Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel.
Shimoni
Eyal
E
Gov
Nir S
NS
Rousso
Itay
I
D000199:Actins / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D001703:Biophysics / Q000379:methods; D003599:Cytoskeleton / Q000378:metabolism; D015683:Gene Products, gag / Q000378:metabolism; D006678:HIV / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D018625:Microscopy, Atomic Force / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D008979:Moloney murine leukemia virus / Q000378:metabolism; D041681:NIH 3T3 Cells; D012190:Retroviridae / Q000378:metabolism*
D000199:Actins; D015683:Gene Products, gag