title: The RNA-binding properties and domain of Rice stripe virus nucleocapsid protein.
abstract: The nucleocapsid protein (NP) of rice stripe virus (RSV) encapsidates viral genomic RNAs to form virion. The binding of NP with RNA is essential for the formation of virus particle. In this study, the binding specificity of RSV NP to RNA and the domains within the NP that mediate this interaction were investigated by gel electrophoretic mobility shift assays and Northwestern blot analysis. The results demonstrated that RSV NP was able to bind to all synthetic RNAs and DNAs without sequence specificity. Using a series of truncated NPs expressed in E. coli and synthetic peptides, we mapped the RNA-binding domain of NP to the central region from amino acid residues 201-232. Further alanine substitution analysis revealed that Lys(206), Lys(207), Lys(220), and Tyr(221) in the RNA-binding domain were essential for NP to bind with RNA.
authors.affiliation: College of Bioscience and Biotechnology, Yangzhou University, No. 12 Wenhui Road East, Yangzhou, 225009, People's Republic of China. slzhao@yzu.edu.cn.
authors.forename: Shuling
authors.initials: S
authors.lastname: Zhao
authors.affiliation: College of Bioscience and Biotechnology, Yangzhou University, No. 12 Wenhui Road East, Yangzhou, 225009, People's Republic of China.
authors.forename: Yanan
authors.initials: Y
authors.lastname: Xue
authors.affiliation: College of Bioscience and Biotechnology, Yangzhou University, No. 12 Wenhui Road East, Yangzhou, 225009, People's Republic of China.
authors.forename: Jiahui
authors.initials: J
authors.lastname: Hao
authors.affiliation: College of Bioscience and Biotechnology, Yangzhou University, No. 12 Wenhui Road East, Yangzhou, 225009, People's Republic of China.
authors.forename: Changyong
authors.initials: C
authors.lastname: Liang
mesh_terms: D015152:Blotting, Northern; D004247:DNA / Q000378:metabolism; D004252:DNA Mutational Analysis; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D016297:Mutagenesis, Site-Directed; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D017384:Sequence Deletion; D013379:Substrate Specificity; D029364:Tenuivirus / Q000235:genetics / Q000502:physiology*
chemical_list: D019590:Nucleocapsid Proteins; D016601:RNA-Binding Proteins; D012313:RNA; D004247:DNA
keywords: Binding domain; EMSA; Nucleocapsid protein; Rice stripe virus