期刊
VETERINARY MICROBIOLOGY
卷 266, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.vetmic.2022.109351
关键词
Fowl Adenovirus serotype 4; Fiber-1 protein; Nuclear localization signal; Importin alpha 1; Co-immunoprecipitation; Co-location
资金
- China Fujian Natural Science Foundation [2019J01658]
- China Fujian Avian Industry System Program [2019-2022]
This study revealed for the first time that the F1 protein of FAdV-4 is located in the nucleus after infection, and confirmed that importin alpha 1 binds to the NLS of F1 protein to localize it in the nucleus, promoting the proliferation of FAdV-4.
Fiber-1 protein (F1) is the structural protein of Fowl Adenovirus serotype 4 (FAdV-4), which could recondite the receptors of host cytomembrane. In this study, we firstly determined that F1 protein located in nucleus of LMH cells after infection with FAdV-4. We additionally revealed that F1 protein had a classic NLS, and the NLS was required for F1 nucleus entry, which was intently associated to the 26th Pro in NLS. The time rule result indi-cated that some F1 proteins firstly positioned in the nucleus 6 h posttranfection, and it entirely located in the nucleus 12 h posttranfection, then it ordinarily placed in cytoplasm 18 h posttranfection by means of microscopic fluorescence observation and Western Blotting. Then after transfection with pCI-neo-flag-F1 or infection with FAdV-4, the importin alpha 1 was once investigated whether or not it was required for F1 protein nucleus entry through immunofluorescence and/or Co-IP, results demonstrated that the F1 protein and importin alpha 1 co-located in the nucleus 6 h and 12 h posttranfection. The tiers of F1 protein vicinity in nucleus have been additionally investigated after knockdown expression or overexpression of importin alpha 1, and the results further revealed that importin alpha 1 used to be required for F1 protein nucleus entry. Finally, the function of F1 protein nucleus entry was investigated by qPCR, RT-PCR and Western Blotting, and the results revealed that F1 protein nucleus location was conducive to the proliferation of FAdV-4. In summary, we firstly reveal that the F1 protein of FAdV-4 locates in nucleus infected with FAdV-4, and confirm that importin alpha 1 binds to the NLS of F1 protein to nucleus localization, which promotes the proliferation of FAdV-4.
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