title: Application of green fluorescent protein-labeled assay for the study of subcellular localization of Newcastle disease virus matrix protein.
abstract: Green fluorescent protein (GFP) used as a powerful marker of gene expression in vivo has so far been applied widely in studying the localizations and functions of protein in living cells. In this study, GFP-labeled assay was used to investigate the subcellular localization of matrix (M) protein of different virulence and genotype Newcastle disease virus (NDV) strains. The M protein of ten NDV strains fused with GFP (GFP-M) all showed nuclear-and-nucleolar localization throughout transfection, whereas that of the other two strains were observed in the nucleus and nucleolus early in transfection but in the cytoplasm late in transfection. In addition, mutations to the previously defined nuclear localization signal in the GFP-M fusion protein were studied as well. Single changes at positions 262 and 263 did not affect nuclear localization of M, while changing both of these arginine residues to asparagine caused re-localization of M mainly to the cytoplasm. The GFP-M was validated as a suitable system for studying the subcellular localization of M protein and could be used to assist us in further identifying the signal sequences responsible for the nucleolar localization and cytoplasmic localization of M protein.
authors.affiliation: College of Veterinary Medicine, Yangzhou University, Yangzhou, Jiangsu, China; Ministry of Education Key Lab for Avian Preventive Medicine, Yangzhou University, Yangzhou, Jiangsu, China.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Duan
authors.forename: Qunhui
authors.initials: Q
authors.lastname: Li
authors.forename: Liang
authors.initials: L
authors.lastname: He
authors.forename: Guo
authors.initials: G
authors.lastname: Zhao
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.forename: Shunlin
authors.initials: S
authors.lastname: Hu
authors.forename: Xiufan
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D002460:Cell Line; D002466:Cell Nucleolus / Q000737:chemistry*; D002467:Cell Nucleus / Q000737:chemistry*; D003593:Cytoplasm / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D006801:Humans; D008856:Microscopy, Fluorescence; D009154:Mutation; D009522:Newcastle disease virus / Q000235:genetics / Q000502:physiology*; D019913:Nuclear Localization Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods*; D014162:Transfection; D014763:Viral Matrix Proteins / Q000032:analysis* / Q000235:genetics
chemical_list: D019913:Nuclear Localization Signals; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; D049452:Green Fluorescent Proteins
keywords: Green fluorescent protein; Matrix protein; Newcastle disease virus; Subcellular localization