Porcine Epidemic Diarrhea Virus ORF3 Protein Is Transported through the Exocytic Pathway.
Amino Acid Sequence
Animals
Coronavirus Infections
/ veterinary
Exocytosis
Genetic Engineering
Host-Pathogen Interactions
Metabolic Networks and Pathways
Open Reading Frames
Plasmids
/ genetics
Porcine epidemic diarrhea virus
/ genetics
Protein Transport
Proteomics
Swine
Swine Diseases
/ metabolism
Viral Proteins
/ chemistry
PEDV
YxxØ
accessory protein
coronavirus
exocytic secretion
intracellular traffic
Journal
Journal of virology
ISSN: 1098-5514
Titre abrégé: J Virol
Pays: United States
ID NLM: 0113724
Informations de publication
Date de publication:
17 08 2020
17 08 2020
Historique:
received:
30
04
2020
accepted:
08
06
2020
pubmed:
20
6
2020
medline:
12
9
2020
entrez:
20
6
2020
Statut:
epublish
Résumé
Accessory genes occurring between the S and E genes of coronaviruses have been studied quite intensively during the last decades. In porcine epidemic diarrhea virus (PEDV), the only gene at this location, ORF3, encodes a 224-residue membrane protein shown to exhibit ion channel activity and to enhance virus production. However, little is known about its intracellular trafficking or about its function during PEDV infection. In this study, two recombinant PEDVs were rescued by targeted RNA recombination, one carrying the full-length ORF3 gene and one from which the gene had been deleted entirely. These viruses as well as a PEDV encoding a naturally truncated ORF3 protein were employed to study the ORF3 protein's subcellular trafficking. In addition, ORF3 expression vectors were constructed to study the protein's independent transport. Our results show that the ORF3 protein uses the exocytic pathway to move to and accumulate in the Golgi area of the cell similarly in infected and transfected cells. Like the S protein, but unlike the other structural proteins M and N, the ORF3 protein was additionally observed at the surface of PEDV-infected cells. In addition, the C-terminally truncated ORF3 protein entered the exocytic pathway but it was unable to leave the endoplasmic reticulum (ER) and ER-to-Golgi intermediate compartment (ERGIC). Consistently, a YxxØ motif essential for ER exit was identified in the C-terminal domain. Finally, despite the use of sensitive antibodies and assays no ORF3 protein could be detected in highly purified PEDV particles, indicating that the protein is not a structural virion component.
Identifiants
pubmed: 32554695
pii: JVI.00808-20
doi: 10.1128/JVI.00808-20
pmc: PMC7431808
pii:
doi:
Substances chimiques
Viral Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
Copyright © 2020 American Society for Microbiology.
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