title: Identification and functional analysis of phosphorylation in Newcastle disease virus phosphoprotein.
abstract: Newcastle disease virus (NDV) encodes a highly phosphorylated P protein; however, the phosphorylation sites have not been identified, and the relationship between phosphorylation and protein function is still unclear. In this study, we bioinformatically predicted 26 amino acid residues in the P protein as potential phosphorylation sites. Furthermore, we treated infected cells with kinase inhibitors to investigate NDV propagation and found that protein kinase C (PKC) is involved in the NDV life cycle and that PKC-activated phosphorylation functions in NDV replication. Using an NDV minigenome assay, we found that expression of a reporter protein decreased when the minigenome system contained P mutants lacking T44, S48, T271, S373 and especially T111. The phosphorylation status of S48, T111, S125 and T271 was determined by Phos-tag SDS-PAGE analysis. Coimmunoprecipitation assays showed that the binding activity of NP and the P-T111A mutant was stronger than that of NP and the wild-type P, suggesting that P-T111 is involved in NP-P interaction. This study sheds light on the mechanism by which P protein phosphorylation affects NDV replication and transcription.
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
authors.forename: Xusheng
authors.initials: X
authors.lastname: Qiu
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhan
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
authors.forename: Chunchun
authors.initials: C
authors.lastname: Meng
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
authors.forename: Junqing
authors.initials: J
authors.lastname: Wang
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
authors.forename: LuNa
authors.initials: L
authors.lastname: Dong
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
authors.forename: Yingjie
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Tan
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
authors.forename: Cuiping
authors.initials: C
authors.lastname: Song
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
authors.forename: Shengqing
authors.initials: S
authors.lastname: Yu
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China. shoveldeen@shvri.ac.cn.|Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, People's Republic of China. shoveldeen@shvri.ac.cn.
authors.forename: Chan
authors.initials: C
authors.lastname: Ding
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D002645:Chickens; D009521:Newcastle Disease / Q000821:virology*; D009522:Newcastle disease virus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011201:Poultry Diseases / Q000821:virology*; D011485:Protein Binding; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D010750:Phosphoproteins; D014764:Viral Proteins; C409209:nucleoprotein, Newcastle disease virus