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Viruses 2014, 6(3), 1253-1273; doi:10.3390/v6031253

Molecular Characterizations of Subcellular Localization Signals in the Nucleocapsid Protein of Porcine Epidemic Diarrhea Virus

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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Author to whom correspondence should be addressed.
Received: 26 November 2013 / Revised: 24 February 2014 / Accepted: 5 March 2014 / Published: 13 March 2014
(This article belongs to the Special Issue Animal Arteriviruses and Coronaviruses)
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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
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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Shi, D.; Lv, M.; Chen, J.; Shi, H.; Zhang, S.; Zhang, X.; Feng, L. Molecular Characterizations of Subcellular Localization Signals in the Nucleocapsid Protein of Porcine Epidemic Diarrhea Virus. Viruses 2014, 6, 1253-1273.

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