title: Localization of an RNA-binding domain in the nucleocapsid protein of the coronavirus mouse hepatitis virus.
abstract: The interaction between the nucleocapsid (N) protein of mouse hepatitis virus (MHV) and RNA was studied in an effort to define portions of the N molecule that participate in binding to RNA. N mRNAs transcribed from SP6 and T7 vectors were translated in a rabbit reticulocyte lysate. Analysis of synthesized N protein in a nondenaturing gel system showed that it bound in vitro to an endogenous RNA in the reticulocyte lysate but not to its own mRNA. A set of deletion mutants was constructed in order to localize the RNA-binding activity of the N protein. It was found that removal of as much as 135 amino-terminal or 57 carboxy-terminal amino acids from the molecule had little or no effect on RNA binding. Moreover, deletion mutants lacking both termini still retained RNA-binding ability. By contrast, internal deletions or truncations extending beyond these two limits effectively abolished RNA binding by N protein. Thus, the RNA-binding region of N has been mapped to the second (central) of the three structural domains of the molecule.
authors.affiliation: Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany.
authors.forename: P S
authors.initials: PS
authors.lastname: Masters
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002213:Capsid / Q000737:chemistry* / Q000378:metabolism; D002474:Cell-Free System; D002872:Chromosome Deletion; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D006517:Murine hepatitis virus / Q000737:chemistry* / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D011817:Rabbits; D015183:Restriction Mapping; D014758:Viral Core Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D012367:RNA, Viral; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins