Guo Shouqing, Liao Yuejiao, Qiu Zhenyu, Liu Geng, Wang Jiamin, Yang Di, Zhang Jiayou, Qiao Zilin, Ma Zhongren, Li Zhuo, Liu Zhenbin
Gansu Tech Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu Univerity, Lanzhou 730030, Gansu, China.
Life Science and Engineering College, Northwest Minzu University, Lanzhou 730030, Gansu, China.
Sheng Wu Gong Cheng Xue Bao. 2022 Mar 25;38(3):1124-1137. doi: 10.13345/j.cjb.210591.
Transglutaminase 2 (TGM2) is a ubiquitous multifunctional protein, which is related to the adhesion of different cells and tumor formation. Previous studies found that TGM2 is involved in the interaction between host cells and viruses, but the effect of TGM2 on the proliferation of influenza virus in cells has not been reported. To explore the effect of TGM2 during H1N1 subtype influenza virus infection, a stable MDCK cell line with TGM2 overexpression and a knockout cell line were constructed. The mRNA and protein expression levels of NP and NS1 as well as the virus titer were measured at 48 hours after pot-infection with H1N1 subtype influenza virus. The results showed that overexpression of TGM2 effectively inhibited the expression of and genes of H1N1 subtype influenza virus, while knockout of TGM2 up-regulated the expression of the and genes, and the expression of the NP at protein level was consistent with that at mRNA level. Virus proliferation curve showed that the titer of H1N1 subtype influenza virus decreased significantly upon TGM2 overexpression. On the contrary, the virus titer in TGM2 knockout cells reached the peak at 48 h, which further proved that TGM2 was involved in the inhibition of H1N1 subtype influenza virus proliferation in MDCK cells. By analyzing the expression of genes downstream of influenza virus response signaling pathway, we found that TGM2 may inhibit the proliferation of H1N1 subtype influenza virus by promoting the activation of JAK-STAT molecular pathway and inhibiting RIG-1 signaling pathway. The above findings are of great significance for revealing the mechanism underlying the interactions between host cells and virus and establishing a genetically engineering cell line for high-yield influenza vaccine production of influenza virus.
转谷氨酰胺酶2(TGM2)是一种广泛存在的多功能蛋白质,与不同细胞的黏附以及肿瘤形成有关。先前的研究发现TGM2参与宿主细胞与病毒之间的相互作用,但TGM2对流感病毒在细胞中增殖的影响尚未见报道。为了探究TGM2在H1N1亚型流感病毒感染过程中的作用,构建了TGM2过表达稳定的MDCK细胞系和敲除细胞系。在接种H1N1亚型流感病毒48小时后,检测NP和NS1的mRNA及蛋白表达水平以及病毒滴度。结果显示TGM2过表达有效抑制了H1N1亚型流感病毒的 和 基因表达,而TGM2敲除则上调了 和 基因的表达,且NP蛋白水平的表达与mRNA水平一致。病毒增殖曲线表明,TGM2过表达时H1N1亚型流感病毒滴度显著下降。相反,TGM2敲除细胞中的病毒滴度在48小时达到峰值,这进一步证明TGM2参与了对MDCK细胞中H1N1亚型流感病毒增殖的抑制。通过分析流感病毒应答信号通路下游基因的表达,我们发现TGM2可能通过促进JAK - STAT分子通路的激活和抑制RIG - 1信号通路来抑制H1N1亚型流感病毒的增殖。上述发现对于揭示宿主细胞与病毒相互作用的机制以及建立用于高产流感病毒疫苗生产的基因工程细胞系具有重要意义。