title: Specific binding of Bluetongue virus NS2 to different viral plus-strand RNAs.
abstract: The Reoviridae have double-stranded RNA genomes of 10-12 segments, each in a single copy in the mature virion. The basis of genome segment sorting during virus assembly that ensures each virus particle contains the complete viral genome is unresolved. Bluetongue virus (BTV) NS2 is a single-stranded RNA-binding protein that forms inclusion bodies in infected cells. Here, we demonstrate that the specific interaction between NS2 and a stem-loop structure present in BTV S10 RNA, and phylogenetically conserved in other BTV serotypes, is abolished by mutations predicted to disrupt the structure. Subsequently, we mapped RNA regions in three other genomic segments of BTV that are bound preferentially by NS2. However, structure probing of these RNAs did not reveal secondary structure motifs that obviously resembled the stem-loop implicated in the NS2-S10 interaction. In addition, the specific binding by NS2 to two different viral RNAs was found to occur independently. Together, these data support the hypothesis that the recognition by NS2 of different RNA structures may be the basis for discrimination between viral RNAs during virus assembly.
authors.affiliation: Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, UK.
authors.forename: Kostas
authors.initials: K
authors.lastname: Lymperopoulos
authors.forename: Rob
authors.initials: R
authors.lastname: Noad
authors.forename: Sara
authors.initials: S
authors.lastname: Tosi
authors.forename: Suran
authors.initials: S
authors.lastname: Nethisinghe
authors.forename: Ian
authors.initials: I
authors.lastname: Brierley
authors.forename: Polly
authors.initials: P
authors.lastname: Roy
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001820:Bluetongue virus / Q000235:genetics / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018411:Spodoptera; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D012367:RNA, Viral; D017361:Viral Nonstructural Proteins