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PMID:23994149
| Citation |
Duan, Z, Li, Q, He, L, Zhao, G, Chen, J, Hu, S and Liu, X (2013) Application of green fluorescent protein-labeled assay for the study of subcellular localization of Newcastle disease virus matrix protein. J. Virol. Methods 194:118-22 |
|---|---|
| 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. |
| Links |
PubMed Online version:10.1016/j.jviromet.2013.08.014 |
| Keywords |
Animals; Cell Line; Cell Nucleolus/chemistry; Cell Nucleus/chemistry; Cytoplasm/chemistry; Green Fluorescent Proteins/analysis; Green Fluorescent Proteins/genetics; Humans; Microscopy, Fluorescence; Mutation; Newcastle disease virus/genetics; Newcastle disease virus/physiology; Nuclear Localization Signals/genetics; Recombinant Fusion Proteins/analysis; Recombinant Fusion Proteins/genetics; Staining and Labeling/methods; Transfection; Viral Matrix Proteins/analysis; Viral Matrix Proteins/genetics |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0044196: host cell nucleolus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleolus. Figure 1A Strain:JS/5/05/Go Accession: JN631747 |
complete | ||||
| GO:0042025: host cell nucleus |
ECO:0000314: |
C |
An green fluorescent protein-labeled assay of GFP-M fusion protein was localized primarily in the nucleus. Figure 1A |
complete | ||||
|
part_of |
GO:0044196: host cell nucleolus |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
| GO:0042025: host cell nucleus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleus. Figure 1B |
complete | ||||
|
Colocalizes with |
GO:0044196: host cell nucleolus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleolus in early infection. Figure 1B Strain:Herts/33 Accession: AY741404 |
complete | |||
|
colocalizes_with |
GO:0044196: host cell nucleolus |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
|
Colocalizes with |
GO:0030430: host cell cytoplasm |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell cytoplasm in late infection. Figure 2 Strain:F48E8 Accession: FJ436302 |
complete | |||
|
colocalizes_with |
GO:0030430: host cell cytoplasm |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
| GO:0042025: host cell nucleus |
ECO:0000314: |
C |
Green fluorescent protein assay of the GFP-M fusion protein was found primarily in the host cell nucleus. Figure 1B |
complete | ||||
| GO:0044196: host cell nucleolus |
ECO:0000314: |
C |
Green fluorescent protein assay of the GFP-M fusion protein was found primarily in the host cell nucleolus. Figure 1B Strain:Mukteswar Accession: ef201805 |
complete | ||||
|
part_of |
GO:0044196: host cell nucleolus |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
| GO:0042025: host cell nucleus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleus. Figure 1B |
complete | ||||
| GO:0044196: host cell nucleolus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleolus. Figure 1B Strain:JS/07/04/Pi Accession: FJ66530 |
complete | ||||
|
part_of |
GO:0044196: host cell nucleolus |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
| GO:0042025: host cell nucleus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleus. Figure 1B |
complete | ||||
| GO:0044196: host cell nucleolus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleolus. Figure 1B Strain:JS/07/22/Pi Accession: FJ66526 |
complete | ||||
|
part_of |
GO:0044196: host cell nucleolus |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
| GO:0042025: host cell nucleus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleus. Figure 1B |
complete | ||||
| GO:0044196: host cell nucleolus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleolus. Figure 1B Strain:V4 Accession: JX524203 |
complete | ||||
|
part_of |
GO:0044196: host cell nucleolus |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
| GO:0042025: host cell nucleus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleus. Figure 1B |
complete | ||||
| GO:0044196: host cell nucleolus |
ECO:0000314: |
C |
Green fluorescent protein assay showed the GFP-M fusion protein was found primarily in the host cell nucleolus. Figure 1B Strain:LaSota Accession: JF950510 |
complete | ||||
|
part_of |
GO:0044196: host cell nucleolus |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
Notes
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