title: Closterovirus bipolar virion: evidence for initiation of assembly by minor coat protein and its restriction to the genomic RNA 5' region.
abstract: The long flexuous virions of the Closteroviridae have a unique bipolar architecture incorporating two coat proteins, with most of the helical nucleocapsid encapsidated by the major coat protein (CP) and a small portion of one end encapsidated by the minor coat protein (CPm). It is not known whether CPm encapsidates the genomic RNA and, if so, which end and what effects transition between the two coat proteins. Two other virus-encoded proteins, an HSP70 homolog (HSP70h) and an approximately 61-kDa protein, are required to augment virion assembly. In this work, we examine the in vivo encapsidation of Citrus tristeza virus by its CPm in the absence of CP. In the absence of other assembly-related proteins, CPm protected a family of 5' coterminal RNAs, apparently because of pausing at different locations along the genomic RNA. Most of the nucleocapsids formed by CPm were short, but a few were full-length and infectious. Mutations within the 5' nontranslated region demonstrated that the CPm origin of assembly overlaps the previously described conserved stem-and-loop structures that function as a cis-acting element required for RNA synthesis. Thus, in the absence of CP, the CPm encapsidation is initiated from the 5' end of the genomic RNA. Coexpression of HSP70h and the p61 protein with CPm in protoplasts restricted encapsidation to the 5' approximately 630 nucleotides, which is close to the normal boundary of the bipolar virion, whereas the presence of either HSP70h or the p61 protein alone did not limit encapsidation by CPm.
authors.affiliation: Department of Plant Pathology, Citrus Research and Education Center, University of Florida, Lake Alfred, FL 33850, USA.
authors.forename: Tatineni
authors.initials: T
authors.lastname: Satyanarayana
authors.forename: Siddarame
authors.initials: S
authors.lastname: Gowda
authors.forename: María A
authors.initials: MA
authors.lastname: Ayllón
authors.forename: William O
authors.initials: WO
authors.lastname: Dawson
mesh_terms: D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics / Q000502:physiology*; D017790:Closterovirus / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure*; D016679:Genome, Viral; D008854:Microscopy, Electron; D009154:Mutation; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics*; D014771:Virion / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure*; D019065:Virus Assembly
chemical_list: D036022:Capsid Proteins; D012367:RNA, Viral