title: Structural and functional analysis of virus factories purified from Rabbit vesivirus-infected Vero cells.
abstract: Rabbit vesivirus infection induces membrane modifications and accumulation of vesicular structures in the cytoplasm of infected Vero cells. Crude RaV replication complexes (RCs) have been purified and their structural and functional properties have been characterized. We show that calnexin, an ER-resident protein, RaV non-structural proteins 2AB-, 2C-, 3A-, 3B- and 3CD-like as well as viral RNAs co-localize within membranous structures which are able to replicate the endogenous RNA templates. The purified virus factories protected their viral RNA contents from microccocal nuclease degradation and were inaccessible to exogenously added synthetic transcripts. In addition, we have shown that RCs can be used to investigate uridylylation of native endogenous VPg. In contrast to the observation that the virus factories were inaccessible to RNAs, RCs were accessible to added recombinant VPg which was subsequently nucleotidylylated. Nevertheless no elongation of an RNA chain attached to native or recombinant VPg could be demonstrated.
authors.affiliation: Servicio Regional de Investigación y Desarrollo Agroalimentario (SERIDA), Laboratorio de Sanidad Animal, Jove del Medio, Gijón, Asturias, Spain.
authors.forename: Rosa
authors.initials: R
authors.lastname: Casais
authors.forename: Lorenzo González
authors.initials: LG
authors.lastname: Molleda
authors.forename: Angeles
authors.initials: A
authors.lastname: Machín
authors.forename: Gloria
authors.initials: G
authors.lastname: del Barrio
authors.forename: Alberto García
authors.initials: AG
authors.lastname: Manso
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Dalton
authors.forename: Ana
authors.initials: A
authors.lastname: Coto
authors.forename: José Manuel Martín
authors.initials: JM
authors.lastname: Alonso
authors.forename: Miguel
authors.initials: M
authors.lastname: Prieto
authors.forename: Francisco
authors.initials: F
authors.lastname: Parra
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000378:metabolism; D002522:Chlorocebus aethiops; D003902:Detergents / Q000494:pharmacology; D019084:Fluorescent Antibody Technique, Indirect; D007668:Kidney / Q000166:cytology* / Q000821:virology*; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D012367:RNA, Viral / Q000378:metabolism; D011817:Rabbits; D014709:Vero Cells; D029342:Vesivirus / Q000187:drug effects / Q000254:growth & development*; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000914:Antibodies, Viral; D003902:Detergents; D012367:RNA, Viral; D014764:Viral Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40