Preliminary study on silencing information regulator 1 restricting influenza A virus replication
10.3760/cma.j.cn112866-20200318-00076
- VernacularTitle:沉默信息调节因子1限制甲型流感病毒复制的初步研究
- Author:
Qiang SUN
1
;
Tingting JIA
;
Shuyi ZHONG
;
Bi HUANG
;
Jiamin CHEN
;
Yuelong SHU
Author Information
1. 中山大学公共卫生学院(深圳)，广州 510000
- Keywords:
Influenza virus;
Silent information regulator 1;
A549 cells;
RNA-Seq;
Differentially expressed genes
- From:
Chinese Journal of Experimental and Clinical Virology
2020;34(3):247-251
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the mechanism of silent information regulator 1 (SIRT1) to restrict influenza virus infection and replication by regulating the downstream proteins.Methods:SIRT1 knockout A549 cell line was constructed by CRISPR-Cas9 technology. The wild-type and SIRT1 knockout A549 cells were infected with influenza A virus. After 24 hours, cells were collected for RNA sequencing (RNA-Seq) and gene set enrichment analysis (GSEA). And the expression levels of the NP protein of influenza virus and SIRT1-related functional proteins in the two cell lines were determined by western blotting.Results:Influenza virus replication was enhanced in SIRT1 knockout A549 cell line. A total of 2 671 differentially expressed genes were detected by high-throughput sequencing, of which 2 012 were up-regulated and 659 were down-regulated, respectively. These differentially expressed genes are involved mainly in signaling pathways such as inflammatory response, apoptosis signal, innate immune antiviral responses, cytokine secretion, and so on. Western blot result indicated that compared with wild-type cell line, the rising degree of the expressions of IFITM3 (interferon-induced transmembrane protein 3) and autophagy protein LC3-II were lower in SIRT1 knockout cell line during influenza virus replication.Conclusions:This study shows that SIRT1 can inhibit influenza virus replication by regulating important proteins downstream such as IFITM3 and LC3-II.
