Cheng Yuqiang, Ma Jingjiao, Liu Yunxia, Gao Quanxin, Yan Yaxian, Wang Hengan, Ding Chan, Sun Jianhe
Shanghai Key Laboratory of Veterinary Biotechnology, Key Laboratory of Urban Agriculture (South), Ministry of Agriculture, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
The Center for Disease Prevention and Control of Baoshan, Shanghai 201901, China.
Dev Comp Immunol. 2017 Dec;77:200-209. doi: 10.1016/j.dci.2017.08.011. Epub 2017 Aug 22.
TANK-binding kinase 1 (TBK1) is an essential serine/threonine-protein kinase required for the Toll-like receptor (TLR)- and retinoic acid-inducible gene I (RIG-I) -mediated induction of type I IFN. Through endogenous Co-IP and LC-MS/MS, we identified chicken TBK1 (chTBK1) as a chSTING-interactive protein. Through exogenous Co-IP assay in transfected cells, we confirmed the interaction between chSTING and chTBK1. To better understand the biological role of chTBK1 in the chSTING-mediated IFN pathway, we cloned the chTBK1 and investigated its biological functions. Quantitative RT-PCR showed that chTBK1 mRNA was widely expressed in different tissues. The overexpression of chTBK1 in DF-1 cells induced the expression of IFN-β and ISGs and inhibited AIV viral replication. We identified indispensable domains of chTBK1 on IFN-β production via the generation of various chTBK1 mutant forms. Together, we identified the chTBK1 as a chSTING interactive protein and concluded that chTBK1 is involved in chSTING-triggered IFN-β signaling in chicken cells.
TANK结合激酶1（TBK1）是Toll样受体（TLR）和视黄酸诱导基因I（RIG-I）介导的I型干扰素诱导所必需的一种重要的丝氨酸/苏氨酸蛋白激酶。通过内源性免疫共沉淀（Co-IP）和液相色谱-串联质谱（LC-MS/MS），我们鉴定出鸡TBK1（chTBK1）是一种与鸡干扰素基因刺激蛋白（chSTING）相互作用的蛋白。通过在转染细胞中进行外源性免疫共沉淀分析，我们证实了chSTING与chTBK1之间的相互作用。为了更好地理解chTBK1在chSTING介导的干扰素途径中的生物学作用，我们克隆了chTBK1并研究了其生物学功能。定量逆转录聚合酶链反应（RT-PCR）表明，chTBK1 mRNA在不同组织中广泛表达。chTBK1在DF-1细胞中的过表达诱导了干扰素-β（IFN-β）和干扰素刺激基因（ISGs）的表达，并抑制了禽流感病毒（AIV）的复制。我们通过构建各种chTBK1突变体形式，确定了chTBK1在IFN-β产生中不可或缺的结构域。总之，我们鉴定出chTBK1是一种与chSTING相互作用的蛋白，并得出结论：chTBK1参与了鸡细胞中chSTING触发的IFN-β信号传导。
