title: Structure and assembly of turnip crinkle virus. VI. Identification of coat protein binding sites on the RNA.
abstract: Structural studies of turnip crinkle virus have been extended to include the identification of high-affinity coat protein binding sites on the RNA genome. Virus was dissociated at elevated pH and ionic strength, and a ribonucleoprotein complex (rp-complex) was isolated by chromatography on Sephacryl S-200. Genomic RNA fragments in the rp-complex, resistant to RNase A and RNase T1 digestion and associated with tightly bound coat protein subunits, were isolated using coat-protein-specific antibodies. The identity of the protected fragments was determined by direct RNA sequencing. These approaches allowed us to study the specific RNA-protein interactions in the rp-complex obtained from dissociated virus particles. The location of one protected fragment downstream from the amber terminator codon in the first and largest of the three viral open reading frames suggests that the coat protein may play a role in the regulation of the expression of the polymerase gene. We have also identified an additional cluster of T1-protected fragments in the region of the coat protein gene that may represent further high-affinity sites involved in assembly recognition.
authors.affiliation: Department of Plant Pathology, University of California, Berkeley 94720.
authors.forename: N
authors.initials: N
authors.lastname: Wei
authors.forename: L A
authors.initials: LA
authors.lastname: Heaton
authors.forename: T J
authors.initials: TJ
authors.lastname: Morris
authors.forename: S C
authors.initials: SC
authors.lastname: Harrison
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D002213:Capsid / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010942:Plant Viruses / Q000502:physiology*; D011485:Protein Binding; D012367:RNA, Viral / Q000378:metabolism*; D014779:Virus Replication / Q000502:physiology*
chemical_list: D012367:RNA, Viral