Subcellular localization of the porcine deltacoronavirus nucleocapsid protein
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ORIGINAL PAPER
Subcellular localization of the porcine deltacoronavirus nucleocapsid protein Zhen Ding1,2 · Suxian Luo1,2 · Wang Gong1,2 · Leyi Wang3 · Nengshui Ding4 · Jun Chen1,2 · Jiajia Chen1,2 · Ting Wang5 · Yu Ye1,2 · Deping Song1,2 · Lingbao Kong5 · Jinghua Zhang1,2 · Yuxing Tang1,2 Received: 15 January 2020 / Accepted: 17 August 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Porcine deltacoronavirus (PDCoV) has been recently identified as an emerging enteropathogenic coronavirus that mainly infects newborn piglets and causes enteritis, diarrhea and high mortality. Although coronavirus N proteins have multifarious activities, the subcellular localization of the PDCoV N protein is still unknown. Here, we produced mouse monoclonal antibodies against the PDCoV N protein. Experiments using anti-haemagglutinin antibodies and these monoclonal antibodies revealed that the PDCoV N protein is shuttled into the nucleolus in both ectopic PDCoV N-expressing cells and PDCoVinfected cells. The results of deletion mutagenesis experiments demonstrated that the predicted nucleolar localization signal at amino acids 295–318 is critical for nucleolar localization. Cumulatively, our study yielded a monoclonal antibody against the PDCoV N protein and revealed a mechanism by which the PDCoV N protein translocated into the nucleolus. The tolls and findings from this work will facilitate further investigations on the functions of the PDCoV N protein. Keywords Porcine deltacoronavirus · Nucleocapsid protein · mAb · Nucleolar localization
Introduction
Edited by Juergen A Richt. * Jinghua Zhang [email protected] * Yuxing Tang [email protected] 1
Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, 1101 Zhimin Dadao, Qingshan Lake District, Nanchang 330045, Jiangxi, China
2
Jiangxi Provincial Key Laboratory for Animal Health, Jiangxi Agricultural University, Nanchang, Jiangxi, China
3
Department of Veterinary Clinical Medicine and the Veterinary Diagnostic Laboratory, College of Veterinary Medicine, University of Illinois, Urbana, IL, USA
4
State Key Laboratory for Pig Genetic Improvement and Production Technology, Jiangxi Agricultural University, Nanchang, Jiangxi, China
5
Institute of Pathogenic Microorganism and College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang, Jiangxi, China
Porcine deltacoronavirus (PDCoV), an emerging swine enteric pathogen, is a major causative agent of watery diarrhea, vomiting, and mortality in piglets, leading to significant losses in the pig industry [1, 2]. In addition to causing global outbreaks of diarrhea in pigs, PDCoV can cross the species barrier to infect calves, chickens, and even humans [3–5], thus posing a significant threat to public health. PDCoV is an enveloped RNA virus with a single-stranded, positive-sense genome. It is classified into the genus Deltacoronavirus, in the family of Coronaviridae and the order Nidovirale.
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