Comprehensive Evolutionary Analysis of Lamprey TNFR-Associated Factors (TRAFs) and Receptor-Interacting Protein Kinase (RIPKs) and Insights Into the Functional Characterization of TRAF3/6 and RIPK1
Comprehensive Evolutionary Analysis of Lamprey TNFR-Associated Factors (TRAFs) and Receptor-Interacting Protein Kinase (RIPKs) and Insights Into the Functional Characterization of TRAF3/6 and RIPK1
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七鳃鳗 TNFR 相关因子 (TRAF) 和受体相互作用蛋白激酶 (RIPK) 的综合进化分析以及对 TRAF3/6 和 RIPK1 功能表征的见解
DOI：
10.3389/fimmu.2020.00663
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发表时间：
2020-04-15
影响因子：
7.3
通讯作者：
Li, Qingwei
中科院分区：
文献类型：
--
作者：
Hou, Jianqiang;Pang, Yue;Li, Qingwei
肿瘤坏死因子相关因子（TRAFs）和受体相互作用蛋白激酶（RIPKs）是哺乳动物体内重要的免疫连接分子，在肿瘤坏死因子α（TNF α）、TLR和IFN信号通路中发挥重要作用。然而，这些基因在脊椎动物中的进化起源以前没有在七鳃鳗中描述过。在这项研究中，我们搜索了七鳃鳗，莱斯纳，和Petromyzon marinus的基因编码的基因，并进行同源序列比对，系统发育树，功能域，保守基序，基因结构和共线性分析，以确定它们的进化关系。七鳃鳗traf和ripk家族的分布和不同病原体刺激七鳃鳗的免疫反应的基因家族也被证明，表明表达和功能多样化的结构变化的作用。此外，双荧光素酶报告基因测定显示，在LjLRIPK 1 a或LjTRAF 3/6过表达中加入外源性免疫调节剂（TNF α或IFN）显著下调NF-κ B或ISRE激活。LjRIPK 1a可显著增强caspase-8活性，并且LjRIPK 1a或LjTRAF 3a/6在HEK 293 T细胞中的过表达导致细胞凋亡。综上所述，本研究对不同脊椎动物中traf和ripk基因家族的理解做出了重要贡献。我们的研究结果也为脊椎动物TRAFs和RIPKs的进化及其对免疫调节的影响提供了新的证据。
TNFR-associated factors (TRAFs) and receptor-interacting protein kinases (RIPKs) are important immunological linker molecules in mammals and play important roles in the TNF alpha, TLR and IFN signaling pathways. However, the evolutionary origins of these genes in vertebrates have not previously been described in lampreys. In this study, we searched the genomes of Lampetra japonicum, Lethenteron reissneri, and Petromyzon marinus for genes encoding trafs and ripks and performed homologous sequence alignment, phylogenetic tree, functional domain, conserved motif, gene structure, and synteny analyses to determine their evolutionary relationships. The distribution of the lamprey traf and ripk families and the immune response of the gene families in lampreys stimulated by different pathogens were also demonstrated, suggesting a role of structural changes in expression and functional diversification. Additionally, the dual luciferase reporter gene assay showed that the addition of exogenous immunomodulator (TNF alpha or IFN) to the overexpression of LjLRIPK1a or LjTRAF3/6 significantly downregulated NF-kappa B or ISRE activation. LjRIPK1a can significantly enhance caspase-8 activity, and overexpression of LjRIPK1a or LjTRAF3a/6 in HEK293T cells results in cell apoptosis. In summary, this study makes an important contribution to the understanding of the traf and ripk gene families in different vertebrates. Our results also provide new evidence for the evolution of vertebrate TRAFs and RIPKs and their impacts on immune regulation.