title: Nucleocapsid Interacts with NPM1 and Protects it from Proteolytic Cleavage, Enhancing Cell Survival, and is Involved in PEDV Growth.
abstract: Porcine epidemic diarrhea virus (PEDV) replicates in the cytoplasm of infected cells, but its nucleocapsid (N) protein localizes specifically to the nucleolus. The mechanism of nuclear translocation, and whether N protein associates with particular nucleolar components, is unknown. In this study, we confirm that a nucleolar phosphoprotein nucleophosmin (NPM1) interacts and co-localizes with the N protein in the nucleolus. In vitro binding studies indicated that aa 148-294 of N and aa 118-188 of NPM1 were required for binding. Interestingly, N protein importation into the nucleolus is independent of the ability of NPM1 to shuttle between the nucleus and the cytoplasm. Furthermore, overexpression of NPM1 promoted PEDV growth, while knockdown of NPM1 suppressed PEDV growth. In addition, binding of N protein to NPM1 protects it from proteolytic degradation by caspase-3, leading to increased cell survival. Taken together, our studies demonstrate a specific interaction of the N protein with the host cell protein NPM1 in the nucleolus. The results suggest potential linkages among viral strategies for the regulation of cell survival activities, possibly through an interaction of N protein with NPM1 which prevents its proteolytic cleavage and enhances cell survival, thus ultimately promoting the replication of PEDV.
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Da
authors.initials: D
authors.lastname: Shi
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Shi
authors.affiliation: College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, No. 2 Xinyang Road, Sartu District, Daqing 163319, P. R. China.
authors.forename: Dongbo
authors.initials: D
authors.lastname: Sun
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Jianfei
authors.initials: J
authors.lastname: Chen
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhang
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Wang
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Jialin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Zhaoyang
authors.initials: Z
authors.lastname: Ji
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Jianbo
authors.initials: J
authors.lastname: Liu
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Liyan
authors.initials: L
authors.lastname: Cao
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Zhu
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yuan
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Dong
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Tiecheng
authors.initials: T
authors.lastname: Chang
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Liu
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin 150069, P. R. China.
authors.forename: Li
authors.initials: L
authors.lastname: Feng
mesh_terms: D021581:Active Transport, Cell Nucleus; D001665:Binding Sites / Q000235:genetics; D002470:Cell Survival; D018352:Coronavirus Infections / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D057809:HEK293 Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D019251:Nucleocapsid / Q000378:metabolism*; D000090243:Nucleophosmin; D053485:Porcine epidemic diarrhea virus / Q000502:physiology*; D011485:Protein Binding; D059748:Proteolysis; D034741:RNA, Small Interfering / Q000235:genetics; D014779:Virus Replication
chemical_list: C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D000090243:Nucleophosmin