Molecular Characterizations of Subcellular Localization Signals in the Nucleocapsid Protein of Porcine Epidemic Diarrhea VirusReportar como inadecuado




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Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China



These authors contributed equally to this work.





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Abstract The nucleolus is a dynamic subnuclear structure, which is crucial to the normal operation of the eukaryotic cell. The porcine epidemic diarrhea virus PEDV, coronavirus nucleocapsid N protein, plays important roles in the process of virus replication and cellular infection. Virus infection and transfection showed that N protein was predominately localized in the cytoplasm, but also found in the nucleolus in Vero E6 cells. Furthermore, by utilizing fusion proteins with green fluorescent protein GFP, deletion mutations or site-directed mutagenesis of PEDV N protein, coupled with live cell imaging and confocal microscopy, it was revealed that, a region spanning amino acids aa, 71–90 in region 1 of the N protein was sufficient for nucleolar localization and R87 and R89 were critical for its function. We also identified two nuclear export signals NES, aa221–236, and 325–364, however, only the nuclear export signal aa325–364 was found to be functional in the context of the full-length N protein. Finally, the activity of this nuclear export signal NES was inhibited by the antibiotic Lepomycin B, suggesting that N is exported by a chromosome region maintenance 1-related export pathway. View Full-Text

Keywords: CHROMOSOME region maintenance 1; nucleocapsid protein; porcine epidemic diarrhea virus; subcellular localization signals CHROMOSOME region maintenance 1; nucleocapsid protein; porcine epidemic diarrhea virus; subcellular localization signals





Autor: Da Shi †, Maojie Lv †, Jianfei Chen, Hongyan Shi, Sha Zhang, Xin Zhang and Li Feng *

Fuente: http://mdpi.com/



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