LncRNA-SUSAJ1 Activates the ER Stress Pathway Inhibiting JEV Proliferation by Promoting PK15 Cells Apoptosis

  • Front Biosci (Landmark Ed). 2022 Sep 13;27(9):260. doi: 10.31083/j.fbl2709260.
Qiongyu Yuan  1 ,  Jinyun Fan  1 ,  Han Wang  1 ,  Xiangchen Li  1 ,  Songbai Yang  1 ,  Ayong Zhao  1 ,  Xiaolong Zhou  1
Affiliations
  • 1. Key Laboratory of Applied Technology on Green Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, 311300 Hangzhou, Zhejiang, China.
PMID: 36224019 DOI: 10.31083/j.fbl2709260
Abstract

Background: Epidemic encephalitis B is a common zoonosis that threatens both pigs and humans. Effective prevention and control of epidemic encephalitis B is difficult. The cellular defence mechanism is closely related to the body's resistance to viral invasion. Long non-coding RNAs (lncRNAs) are involved in regulating various cellular activities. We previously found that lncRNA-SUSAJ1 could inhibit the proliferation of Japanese encephalitis virus (JEV). However, the mechanism underlying this suppression remains unclear.

Methods: We performed Western blotting and quantitative reverse-transcription polymerase chain reaction (RT-qPCR) analyses, as well as mitochondrial membrane potential, flow cytometry, terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL), RNA pull-down, and RNA immunoprecipitation assays.

Results: JC-1 cationic dye staining showed that lncRNA-SUSAJ1 promoted the depolarisation of mitochondrial membrane potential; H2DCFDA probe staining showed that lncRNA-SUSAJ1 enhanced the level of Reactive Oxygen Species in PK15 porcine kidney cells. qRT-PCR and Western blotting revealed the expression levels of associated mRNAs and proteins, and the TUNEL and flow cytometry assays detected cell Apoptosis. Their results showed that lncRNA-SUSAJ1 promoted the expression of pro-apoptotic genes and inhibited the expression of anti-apoptotic genes. RNA pull-down experiments using biotin-labelled lncRNA-SUSAJ1 showed colocalisation between lncRNA-SUSAJ1 and the 70 kDa heat shock protein (HSP70). lncRNA-SUSAJ1 also activated unfolded protein response-related pathways, regulated protein degradation, and promoted Apoptosis via the endoplasmic reticulum stress response, thereby inhibiting viral replication.

Conclusions: The findings of this study provide insight into the specific molecular mechanism of lncRNA-SUSAJ1 resistance to viral proliferation by promoting cell Apoptosis, clarify the Antiviral effect of lncRNA-SUSAJ1 on JEV.

Keywords
JEV; apoptosis; endoplasmic reticulum stress response; lncRNA-SUSAJ1.
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