Subcellular localization of the porcine deltacoronavirus nucleocapsid protein

Virus Genes. 2020 Dec;56(6):687-695. doi: 10.1007/s11262-020-01790-0. Epub 2020 Sep 17.

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 PDCoV-infected 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: Nucleocapsid protein; Nucleolar localization; Porcine deltacoronavirus; mAb.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Viral / biosynthesis
  • Antibodies, Viral / chemistry
  • Cell Line
  • Cell Nucleolus / genetics*
  • Cell Nucleolus / metabolism
  • Coronavirus Infections / pathology
  • Coronavirus Infections / virology*
  • Coronavirus Nucleocapsid Proteins / genetics*
  • Coronavirus Nucleocapsid Proteins / metabolism
  • Deltacoronavirus / genetics*
  • Deltacoronavirus / growth & development
  • Deltacoronavirus / metabolism
  • Epithelial Cells / metabolism
  • Epithelial Cells / ultrastructure
  • Epithelial Cells / virology
  • Gastroenteritis, Transmissible, of Swine / pathology
  • Gastroenteritis, Transmissible, of Swine / virology*
  • Gene Expression
  • Hemagglutinins, Viral / genetics
  • Hemagglutinins, Viral / metabolism
  • Host-Pathogen Interactions / genetics*
  • Kidney / pathology
  • Kidney / virology
  • Mice
  • Nuclear Localization Signals
  • Protein Transport
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sequence Deletion
  • Swine

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Coronavirus Nucleocapsid Proteins
  • Hemagglutinins, Viral
  • Nuclear Localization Signals
  • Recombinant Proteins