title: Molecular characterizations of subcellular localization signals in the nucleocapsid protein of porcine epidemic diarrhea virus.
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.
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. dashi198566@163.com.
authors.forename: Da
authors.initials: D
authors.lastname: Shi
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. lvmaojie650@sina.com.
authors.forename: Maojie
authors.initials: M
authors.lastname: Lv
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. chenjianfei@126.com.
authors.forename: Jianfei
authors.initials: J
authors.lastname: Chen
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. shy2005y@163.com.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Shi
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. zs8838566@126.com.
authors.forename: Sha
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. zhangxin2410@163.com.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. fl@hvri.ac.cn.
authors.forename: Li
authors.initials: L
authors.lastname: Feng
mesh_terms: D000818:Animals; D002466:Cell Nucleolus / Q000737:chemistry; D002522:Chlorocebus aethiops; D000086462:Coronavirus Nucleocapsid Proteins; D003593:Cytoplasm / Q000737:chemistry; D004252:DNA Mutational Analysis; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000378:metabolism; D053485:Porcine epidemic diarrhea virus / Q000235:genetics* / Q000502:physiology; D021382:Protein Sorting Signals; D014709:Vero Cells
chemical_list: D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins; D021382:Protein Sorting Signals