Black carp LGP2 suppresses RIG-I mediated IFN signaling during the antiviral innate immunity.
Black carp LGP2 suppresses RIG-I mediated IFN signaling during the antiviral innate immunity.
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DOI：
10.1016/j.fsi.2023.109208
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发表时间：
2023-11
影响因子：
4.7
通讯作者：
Yixuan He;Ji Liu;Yujia Miao;Meiling Liu;Hui-na Wu;Jun Xiao;Hao Feng
中科院分区：
文献类型：
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作者：
Yixuan He;Ji Liu;Yujia Miao;Meiling Liu;Hui-na Wu;Jun Xiao;Hao Feng
遗传学和生理学实验室2（LGP 2）是视黄酸诱导基因（RIG）-I样受体（RLR）的成员，在哺乳动物和硬骨鱼类中均参与IFN信号转导。在我们以前的研究中，青鱼（Mylopharyngodon piceus）LGP 2（bcLGP 2）已被鉴定为正调控黑色素瘤分化相关基因5（MDA 5）。在本研究中，bcLGP 2的敲低降低了宿主基因的表达，包括bcIFNb，bcPKR，bcMx 1和bcViperin，也削弱了宿主细胞的抗病毒能力。探讨了bcLGP 2与青鱼RIG-Ib（bcRIG-Ib）的关系。双荧光素酶报告基因检测和qRT-PCR检测表明，bcLGP 2抑制了bcRIG-Ib诱导的I型干扰素（IFN）和干扰素刺激基因（ISG）的转录，包括PKR、ISG 15和Viperin。空斑试验证实bcLGP 2减弱了bcRIG-Ib介导的抗鲤鱼春季病毒血症病毒（SVCV）的能力。免疫共沉淀法鉴定了bcLGP 2与bcRIG-Ib以及bcLGP 2与bcRIG-Ib-CARD之间的相互作用。bcLGP 2也减弱了bcRIG-Ib-CARD介导的抗病毒能力。截断突变分析表明，bcLGP 2的DExD/H-box解旋酶结构域对bcRIG-Ib的抑制作用与bcLGP 2相似，而C端阻遏结构域（CTD）对bcRIG-Ib的抑制作用不明显。此外，bcLGP 2增强了bcRIG-Ib的K48连接的泛素化，促进了bcRIG-Ib的蛋白酶体依赖性降解。因此，我们的数据支持这样的结论，即bcLGP 2通过蛋白酶体与bcRIG-Ib相互作用并诱导其降解，从而抑制了bcRIG-Ib介导的抗病毒信号传导。
Laboratory of genetics and physiology 2 (LGP2), a member of retinoic acid-inducible gene (RIG)-I-like receptors (RLRs), has been reported to play different roles in IFN signaling in both mammals and teleost fish. In our previous study, black carp (Mylopharyngodon piceus) LGP2 (bcLGP2) has been characterized to positively regulate melanoma differentiation-associated gene 5 (MDA5). In this study, knockdown of bcLGP2 decreased the expression of host genes, includingbcIFNb,bcPKR,bcMx1, andbcViperin, and also attenuated the antiviral capability of host cells. The relationship between bcLGP2 and black carp RIG-Ib (bcRIG-Ib) has been explored. Dual-luciferase reporter assay and qRT-PCR assay indicated that bcLGP2 dampened bcRIG-Ib induced transcription of type I interferons (IFNs) and interferon-stimulated genes (ISGs), includingPKR,ISG15, andViperin. Consistently, the plaque assay identified that bcLGP2 attenuated bcRIG-Ib mediated antiviral ability against spring viremia of carp virus (SVCV). Co-immunoprecipitation assay identified the interaction between bcLGP2 and bcRIG-Ib, as well as bcLGP2 and bcRIG-Ib-CARD. And bcRIG-Ib-CARD mediated antiviral ability was also attenuated by bcLGP2. Truncation mutation analysis showed DExD/H-box Helicase domain of bcLGP2 possessed a similar inhibitory effect on bcRIG-Ib to that of bcLGP2, while the C-terminus repressor domain (CTD) presented little impact on bcRIG-Ib. Furthermore, bcLGP2 enhanced the K48-linked ubiquitination of bcRIG-Ib, promoting proteasome-dependent degradation of bcRIG-Ib. Thus, our data supported the conclusion that bcLGP2 interacted with and induced degradation of bcRIG-Ib through proteasome, leading to the dampened antiviral signaling mediated by bcRIG-Ib.