Modulation of Porcine Epidemic Diarrhea Virus (PEDV) RNA translation by the nucleocapsid protein
| dc.contributor.committeeMember | Tikoo, Suresh | |
| dc.contributor.committeeMember | Falzarano, Darryl | |
| dc.contributor.committeeMember | Luo, Yu | |
| dc.creator | Hao, Lin | |
| dc.creator.orcid | 0009-0005-4071-4037 | |
| dc.date.accessioned | 2024-09-03T15:24:23Z | |
| dc.date.available | 2024-09-03T15:24:23Z | |
| dc.date.copyright | 2024 | |
| dc.date.created | 2024-07 | |
| dc.date.issued | 2024-08-29 | |
| dc.date.submitted | July 2024 | |
| dc.date.updated | 2024-09-03T15:24:23Z | |
| dc.description.abstract | Porcine epidemic diarrhea (PED) is a serious epidemic outbreak, characterized by vomiting, diarrhea, dehydration, and anorexia in pigs of all ages. It brings problems to pig industry and causes significant economic losses. However, according to the effectiveness of current porcine epidemic diarrhea virus (PEDV) vaccines is not very high, so it is very important to develop new and effective PEDV vaccines and therapeutics. The PEDV nucleocapsid (N) protein is a highly conserved protein, which usually be phosphorylated. It has multiple functions. For example, as a structural protein, it plays a role in the nucleocapsid formation with viral genomic RNA. Moreover, it regulates viral replication, transcription, and assembly. At the same time, the N-terminal domain and serine arginine-rich (SR) region of some coronavirus N proteins are usually modified by phosphorylation, which contributes to increasing RNA binding and is essential for replication. In addition, the C-terminal domain of N protein mediates dimerization. Although it has been shown that N protein plays an important role in both virus RNA synthesis and host cell processes regulation, the effect of PEDV N protein on viral translation is not well understood. We therefore studied the role of PEDV N protein in regulating viral RNA translation. Our results showed that N protein increases PEDV RNA translation. Furthermore, we discovered a synergistic impact of the N-terminal domain (NTD) and the linker region in increasing PEDV translation. Additionally, our research demonstrated that N protein dramatically increases PEDV RNA translation when only 3' untranslated region is present. Moreover, PEDV N protein regulates viral RNA translation through the 3' untranslated region bulged stem loop (BSL) domain. Akt is a serine/threonine–protein kinase which exists as three isoforms: Akt1, Akt2 and Akt3. Both Akt1 and Akt2 have been found to be activated by virus infections and play a role in regulating viral replication or translation. However, whether one or more Akt isoforms influence PEDV translation remains unclear. We performed ectopic expression and knockdown experiments to investigate the effects of Akt isoforms on viral RNA translation modulation by PEDV N protein. Results showed that Akt1 increases the enhancement of RNA translation by the N protein. Furthermore, we demonstrated that Akt1 enhancement of viral translation depends on its kinase activity and catalytic domain. We provided evidence of the interaction between PEDV N and Akt1 through GST pull-down and co-localization assays. Moreover, we revealed that the NTD and Linker region of PEDV N protein decreases its interaction with Akt1 but increases PEDV RNA translation enhancement by Akt1. In summary, this study indicated PEDV N protein increases RNA translation and Akt1 enhances viral translation upregulation by N protein. | |
| dc.format.mimetype | application/pdf | |
| dc.identifier.uri | https://hdl.handle.net/10388/15965 | |
| dc.language.iso | en | |
| dc.subject | Porcine epidemic diarrhea virus (PEDV), nucleocapsid protein, PEDV RNA translation, Akt | |
| dc.title | Modulation of Porcine Epidemic Diarrhea Virus (PEDV) RNA translation by the nucleocapsid protein | |
| dc.type | Thesis | |
| dc.type.material | text | |
| thesis.degree.department | School of Public Health | |
| thesis.degree.discipline | Vaccinology and Immunotherapeutics | |
| thesis.degree.grantor | University of Saskatchewan | |
| thesis.degree.level | Masters | |
| thesis.degree.name | Master of Science (M.Sc.) |