Importin α5 negatively regulates importin β1-mediated nuclear import of Newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblasts
The matrix (M) protein of Newcastle disease virus (NDV) is demonstrated to localize in the nucleus via intrinsic nuclear localization signal (NLS), but cellular proteins involved in the nuclear import of NDV M protein and the role of M's nuclear localization in the replication and pathogenicity...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2018-12-01
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| Series: | Virulence |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/21505594.2018.1449507 |
| _version_ | 1811321813114814464 |
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| author | Zhiqiang Duan Haixu Xu Xinqin Ji Jiafu Zhao Houqiang Xu Yan Hu Shanshan Deng Shunlin Hu Xiufan Liu |
| author_facet | Zhiqiang Duan Haixu Xu Xinqin Ji Jiafu Zhao Houqiang Xu Yan Hu Shanshan Deng Shunlin Hu Xiufan Liu |
| author_sort | Zhiqiang Duan |
| collection | DOAJ |
| description | The matrix (M) protein of Newcastle disease virus (NDV) is demonstrated to localize in the nucleus via intrinsic nuclear localization signal (NLS), but cellular proteins involved in the nuclear import of NDV M protein and the role of M's nuclear localization in the replication and pathogenicity of NDV remain unclear. In this study, importin β1 was screened to interact with NDV M protein by yeast two-hybrid screening. This interaction was subsequently confirmed by co-immunoprecipitation and pull-down assays. In vitro binding studies indicated that the NLS region of M protein and the amino acids 336–433 of importin β1 that belonged to the RanGTP binding region were important for binding. Importantly, a recombinant virus with M/NLS mutation resulted in a pathotype change of NDV and attenuated viral replication and pathogenicity in chicken fibroblasts and SPF chickens. In agreement with the binding data, nuclear import of NDV M protein in digitonin-permeabilized HeLa cells required both importin β1 and RanGTP. Interestingly, importin α5 was verified to interact with M protein through binding importin β1. However, importin β1 or importin α5 depletion by siRNA resulted in different results, which showed the obviously cytoplasmic or nuclear accumulation of M protein and the remarkably decreased or increased replication ability and pathogenicity of NDV in chicken fibroblasts, respectively. Our findings therefore demonstrate for the first time the nuclear import mechanism of NDV M protein and the negative regulation role of importin α5 in importin β1-mediated nuclear import of M protein and the replication and pathogenicity of a paramyxovirus. |
| first_indexed | 2024-04-13T13:24:21Z |
| format | Article |
| id | doaj.art-e15db8020f4c4bafae0d9ca0a703f708 |
| institution | Directory Open Access Journal |
| issn | 2150-5594 2150-5608 |
| language | English |
| last_indexed | 2024-04-13T13:24:21Z |
| publishDate | 2018-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Virulence |
| spelling | doaj.art-e15db8020f4c4bafae0d9ca0a703f7082022-12-22T02:45:13ZengTaylor & Francis GroupVirulence2150-55942150-56082018-12-019178380310.1080/21505594.2018.14495071449507Importin α5 negatively regulates importin β1-mediated nuclear import of Newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblastsZhiqiang Duan0Haixu Xu1Xinqin Ji2Jiafu Zhao3Houqiang Xu4Yan Hu5Shanshan Deng6Shunlin Hu7Xiufan Liu8Guizhou UniversityYangzhou UniversityGuizhou UniversityGuizhou UniversityGuizhou UniversityGuizhou UniversityGuizhou UniversityYangzhou UniversityYangzhou UniversityThe matrix (M) protein of Newcastle disease virus (NDV) is demonstrated to localize in the nucleus via intrinsic nuclear localization signal (NLS), but cellular proteins involved in the nuclear import of NDV M protein and the role of M's nuclear localization in the replication and pathogenicity of NDV remain unclear. In this study, importin β1 was screened to interact with NDV M protein by yeast two-hybrid screening. This interaction was subsequently confirmed by co-immunoprecipitation and pull-down assays. In vitro binding studies indicated that the NLS region of M protein and the amino acids 336–433 of importin β1 that belonged to the RanGTP binding region were important for binding. Importantly, a recombinant virus with M/NLS mutation resulted in a pathotype change of NDV and attenuated viral replication and pathogenicity in chicken fibroblasts and SPF chickens. In agreement with the binding data, nuclear import of NDV M protein in digitonin-permeabilized HeLa cells required both importin β1 and RanGTP. Interestingly, importin α5 was verified to interact with M protein through binding importin β1. However, importin β1 or importin α5 depletion by siRNA resulted in different results, which showed the obviously cytoplasmic or nuclear accumulation of M protein and the remarkably decreased or increased replication ability and pathogenicity of NDV in chicken fibroblasts, respectively. Our findings therefore demonstrate for the first time the nuclear import mechanism of NDV M protein and the negative regulation role of importin α5 in importin β1-mediated nuclear import of M protein and the replication and pathogenicity of a paramyxovirus.http://dx.doi.org/10.1080/21505594.2018.1449507Newcastle disease virusmatrix proteinnuclear localization signalnuclear import mechanismchicken fibroblasts |
| spellingShingle | Zhiqiang Duan Haixu Xu Xinqin Ji Jiafu Zhao Houqiang Xu Yan Hu Shanshan Deng Shunlin Hu Xiufan Liu Importin α5 negatively regulates importin β1-mediated nuclear import of Newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblasts Virulence Newcastle disease virus matrix protein nuclear localization signal nuclear import mechanism chicken fibroblasts |
| title | Importin α5 negatively regulates importin β1-mediated nuclear import of Newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblasts |
| title_full | Importin α5 negatively regulates importin β1-mediated nuclear import of Newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblasts |
| title_fullStr | Importin α5 negatively regulates importin β1-mediated nuclear import of Newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblasts |
| title_full_unstemmed | Importin α5 negatively regulates importin β1-mediated nuclear import of Newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblasts |
| title_short | Importin α5 negatively regulates importin β1-mediated nuclear import of Newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblasts |
| title_sort | importin α5 negatively regulates importin β1 mediated nuclear import of newcastle disease virus matrix protein and viral replication and pathogenicity in chicken fibroblasts |
| topic | Newcastle disease virus matrix protein nuclear localization signal nuclear import mechanism chicken fibroblasts |
| url | http://dx.doi.org/10.1080/21505594.2018.1449507 |
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