title: Characterisation of interaction between NS3 and NS5B protein of classical swine fever virus by deletion of terminal sequences of NS5B.
abstract: The NS3-NS5B interaction of classical swine fever virus (CSFV) is important for viral replication. For characterisation of the interaction between the NS3 and NS5B, a series of NS5B mutants with deletion of N-, C-terminal amino acids and quadruple alanine substitution mutations were produced. GST pull-down assays and immunoprecipitation analyses showed that NS5B and some NS5B mutants have NS3 binding activity. Further experimental data indicated that CSFV NS5B might contain two NS3 binding sites, one covering amino acids 63-99 located at the N-terminal end, another covering amino acids 611-642 at the C-terminal end. Assays for RNA-dependent RNA polymerase (RdRp) activity revealed that CSFV NS3 is able to enhance the RdRp activity of NS5B and some NS5B mutants in vitro. The enhancement might be obtained by NS3 binding to the two terminal sequences of NS5B, which could be attractive targets for drug development against CSFV.
authors.affiliation: College of Life and Environment Sciences, Shanghai Normal University, Shanghai, 200234, China.
authors.forename: Yujing
authors.initials: Y
authors.lastname: Wang
authors.forename: Zailing
authors.initials: Z
authors.lastname: Zhu
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.forename: Jialin
authors.initials: J
authors.lastname: Yu
authors.forename: Lingzhu
authors.initials: L
authors.lastname: Wan
authors.forename: Jun
authors.initials: J
authors.lastname: Chen
authors.forename: Ming
authors.initials: M
authors.lastname: Xiao
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D006692:Classical Swine Fever Virus / Q000235:genetics* / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics*; D013552:Swine; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
chemical_list: D017361:Viral Nonstructural Proteins; D012324:RNA-Dependent RNA Polymerase