title: [Matrix protein of Newcastle disease virus interacts with avian nucleophosmin B23. 1 in HEK-293T cells].
abstract: OBJECIVE: To identify the interaction between Newcastle disease virus (NDV) matrix (M) protein and avian nucleophosmin B23. 1 in HEK-293T cells.
  
  
  METHODS
  Specific primers used to amplify M gene and B23. 1 gene were designed and synthesized according to JS/5/05/Go whole gene sequence (JN631747) and avian nucleophosmin B23. 1 gene sequence (NM_205267). Viral RNA and cellular RNA were extracted from allantoic fluid of NDV JS/5/05/Go strain and DF1 cells with TRIzol reagent, respectively. The M gene and B23. 1 gene were amplified by RT-PCR and then subcloned into eukaryotic expression vectors to generate the recombinant plasmids pEGFP-M, pCMV-HA-M and pDsRed-B23. 1. To investigate the localization features of M protein and B23. 1 protein, we transfected the plasmids pEGFP-M and pDsRed-B23. 1 simultaneously into HEK-293T cells and observed the results by fluorescence microscopy. We further confirmed the interaction between the two proteins by co-immunoprecipitation (Co-IP) assays.
  
  
  RESULTS
  The fusion proteins were successfully expressed in transfected HEK-293T cells by Western blot analysis. The NDV M protein and avian nucleophosmin B23. 1 showed co-localization features in the nucleolus in co-transfected HEK-293T cells. Furthermore, the binding of M with B23. 1 was demonstrated by Co-IP assays.
  
  
  CONCLUSIONS
  These results suggested that NDV M protein interacted with avian nucleophosmin B23. 1 and the nucleolar localization of M might be regulated via interaction with B23. 1.
authors.affiliation: Ministry of Education Key Lab for Avian Preventive Medicine, Yangzhou University, Yangzhou 225009, China. dzq2040847@yahoo.com.cn
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Duan
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.forename: Liang
authors.initials: L
authors.lastname: He
authors.forename: Haixu
authors.initials: H
authors.lastname: Xu
authors.forename: Qunhui
authors.initials: Q
authors.lastname: Li
authors.forename: Shunlin
authors.initials: S
authors.lastname: Hu
authors.forename: Xiufan
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D002645:Chickens; D057809:HEK293 Cells; D006801:Humans; D009521:Newcastle Disease / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D009522:Newcastle disease virus / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D000090243:Nucleophosmin; D011485:Protein Binding; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D014763:Viral Matrix Proteins; D000090243:Nucleophosmin