Grass carp ATG5 and ATG12 promote autophagy but down-regulate the transcriptional expression levels of IFN-I signaling pathway
Grass carp ATG5 and ATG12 promote autophagy but down-regulate the transcriptional expression levels of IFN-I signaling pathway
复制标题
草鱼ATG5和ATG12促进自噬但下调IFN-I信号通路转录表达水平
DOI：
10.1016/j.fsi.2019.06.014
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发表时间：
2019-09-01
影响因子：
4.7
通讯作者：
Wang, Yaping
中科院分区：
文献类型：
--
作者：
Chu, Pengfei;He, Libo;Wang, Yaping
自噬是一个基本和保守的过程，在生理动态平衡、应激适应性反应和免疫反应中发挥着重要作用。自噬相关蛋白(ATGs)是自噬机制的关键组成部分。本研究鉴定了草鱼自噬相关基因5(ATG5)和12(ATG12)。在鳃和肠道中，ATG5和ATG12高表达，但在草鱼呼肠孤病毒(GCRV)感染后，它们的表达显著降低。在草鱼肾(CIK)细胞中，观察到Poly(I：C)感染后ATG5和ATG12表达的急剧变化。亚细胞定位显示ATG5和ATG12均匀分布于胞浆和胞核。而ATG5和ATG12的相互作用仅见于293T细胞和CIK细胞的胞浆中。此外，ATG5和ATG12在293T细胞中的过表达表现为自噬增强，当ATG5和ATG12同时过表达时，自噬过程更容易进行。双荧光素酶活性检测表明，ATG5和ATG12均能显著抑制IRF3、IRF7和干扰素-I的启动子活性。此外，ATG5和ATG12结合物对Poly(I：C)或GCRV感染的干扰素-I表达的抑制作用远强于ATG5或ATG12。综上所述，这些结果表明，草鱼ATG5和ATG12在先天免疫和自噬中起着重要作用。
Autophagy is an essential and conserved process that plays an important role in physiological homeostasis, adaptive response to stress and the immune response. Autophagy-related proteins (ATGs) are key components of the autophagic machinery. In the study, grass carp (Ctenophcayngodon idella) autophagy-related gene 5 (ATG5) and 12 (ATG12) were identified. In the gill and intestine, ATG5 and ATG12 were highly expressed, but after grass carp reovirus (GCRV) infection, they were decreased significantly. In Ctenopharyrtgodon idella kidney (CIK) cells, the sharp variation of ATG5 and ATG12 expression was observed after poly(I:C) infection. Subcellular localisation showed that ATG5 and ATG12 were evenly distributed in the cytoplasm and nucleus. However, the interaction between ATG5 and ATG12 was only found in cytoplasm in both 293T cells and CIK cells. In addition, the overexpression of ATG5 or ATG12 in 293T cells showed enhanced autophagy, and autophagic process was facilitated when ATG5 and ATG12 were simultaneously overexpressed. Dual-luciferase activity assay indicated that both ATG5 and ATG12 remarkably suppressed the promoter activity of IRF3, IRF7, and IFN-I. Further, ATG5 and ATG12 conjugate showed far stronger inhibitory affection on the expression of IFN-I than either ATG5 or ATG12 in response to poly(I:C) or GCRV infection. Taken together, the results demonstrate that grass carp ATG5 and ATG12 play an important role in innate immunity and autophagy.