N 6-methyladenosine modification contributes to respiratory syncytial virus infection

Zhu Li; Yi Liu; Ling Zhang; Jiahua Tian; Hongping Wang; Hongwei Ding; Jin Nie; Hang Pi; Bingyao Wang; Daishun Liu

doi:10.1016/j.heliyon.2023.e15307

N ⁶-methyladenosine modification contributes to respiratory syncytial virus infection

Heliyon. 2023 Apr 6;9(4):e15307. doi: 10.1016/j.heliyon.2023.e15307. eCollection 2023 Apr.

Authors

Zhu Li^{1

2}, Yi Liu³, Ling Zhang^{2

3}, Jiahua Tian⁴, Hongping Wang⁵, Hongwei Ding⁵, Jin Nie⁴, Hang Pi³, Bingyao Wang³, Daishun Liu^{1

5}

Affiliations

¹ Medical College of Soochow University, 215000, Suzhou, China.
² Department of Respiratory Medicine, The First People's Hospital of Zunyi, (The Third Affiliated Hospital of Zunyi Medical University), 563000, Guizhou, China.
³ Zhuhai Campus of Zunyi Medical University, Zhuhai, 519000, Guangdong, China.
⁴ The Affiliated Hospital of Zunyi Medical University, 563000, Guizhou, China.
⁵ Zunyi Medical University, Zunyi, 563000, Guizhou, China.

Abstract

Background: Respiratory syncytial virus (RSV) is the second leading cause of death due to lower respiratory tract infections. Effective prevention and treatment measures are lacking, posing a huge socioeconomic burden to the world. N ⁶-methyladenosine (m⁶A) is the most common internal modification in messenger RNA and noncoding RNA. Numerous recent studies have shown that the dysregulation of m⁶A modification is associated with diseases caused by pathogenic viruses.

Methods: The changes in m⁶A modification were evaluated using m⁶A RNA methylation assay. The differences in gene expression levels of various m⁶A-modifying enzymes were observed using Quantitative Real-time PCR (qRT-PCR) during RSV infection. The autophagosomes were observed using transmission electron microscopy, and the expression of autophagy-associated protein Microtubule Associated Protein 1 Light Chain 3 Beta Ⅱ/Ⅰ (LC3B Ⅱ/Ⅰ) and Beclin1 in Human Normal Lung Epithelial Cells (BEAS-2B) cells using Western blot during RSV infection. The significantly differentially expressed genes were screened guided by bioinformatics. Their relationship with m⁶A-modifying enzymes was analyzed through protein-protein interaction network and expression correlation analysis.

Results: The m⁶A abundance decreased and demethylase Fat Mass and Obesity- Associated Protein (FTO) significantly increased during RSV infection after 24 h. We also found that the DNA Damage-Inducible Transcript 3 Protein (DDIT3) level significantly increased during RSV infection after 24 h and observed autophagosomes in BEAS-2B cells. In addition, RSV infection could cause the upregulation of LC3B Ⅱ/Ⅰ and Beclin1. The expression correlation analysis showed that DDIT3 levels were positively correlated with the FTO level, and Methyltransferase Like 3 (METTL3), RNA Binding Motif Protein 15B (RBM15B), YTH Domain-Containing Family Protein 1 (YTHDF1), and levels were negatively correlated with the DDIT3 level.

Conclusions: We uncovered a significant role for m⁶A modification during RSV infection. Also, a correlation was found between m⁶A and autophagy, providing new ideas for therapeutic advancements in RSV treatment.

Keywords: Autophagy; Bioinformatics; Interaction; N6-methyladenosine; RSV.