National and Provincial Joint Laboratory of Exploration and Utilization of Marine Aquatic Genetic Resources, National Engineering Research Center of Marine Facilities Aquaculture, Zhejiang Ocean University, Zhoushan, 316022, PR China.
Ningbo International Travel Health Care Center, Ningbo, 315012, PR China.
Fish Shellfish Immunol. 2020 Jul;102:469-479. doi: 10.1016/j.fsi.2020.05.006. Epub 2020 May 8.
Interleukin-1 receptor-associated kinases (IRAKs) play important roles in the innate immune system of TLR (Toll-like receptor) signaling pathway. In this paper, interleukin-1 receptor-associated kinase-b (designated as McIRAK-b) and interleukin-1 receptor-associated kinase-a (named as McIRAK-a) were obtained based on the transcriptome data, the full length of McIRAK-b was 1815 bp and McIRAK-a was 3168bp, encoding 532 and 978 amino acids, respectively. BLASTp analysis and phylogenetic relationship strongly suggested that the deduced amino acid sequence of McIRAK-b had high homology with IRAK-4 and McIRAK-a was similar to IRAK-1 of other mollusks, especially at their function domains. The expressions of McIRAK-b and McIRAK-a were detected in six tissues including adductor muscle, hemocyte, gills, gonad and hepatopancreas, and the highest expressions appeared both in gills. The expressions of McIRAK-b and McIRAK-a in gills were observed with time-dependent manners after bacterial infections. After being challenged with Vibrio alginolyticus, McIRAK-b expressed significantly and got the peak at 8 h (9.47 times compared with the control group), but the peak appeared at 4 h by being infected with Vibrio parahaemolyticus (12.02 times compared with the control group). The highest point of McIRAK-a mRNA appeared at 12 h (5.16 times) after being challenged with V.alginolyticus and 8 h (4.21 times) for V.parahaemolyticus challenge. The results suggested that IRAK-b and IRAK-a might be important in immune signaling pathway of mussels. The kinase functional domain sequences (S_TKc) of McIRAK-b and McIRAK-a expressed in BL21(DE3) and purified by Ni-NAT Superflow resin conforming to the expected molecular weight with many active sites for their conferring protein-protein interaction functions. This study may provide some further understandings of the regulatory mechanisms in the bivalve innate immune system for IRAKs family.
白细胞介素-1 受体相关激酶 (IRAKs) 在 TLR (Toll 样受体) 信号通路的固有免疫系统中发挥重要作用。在本文中，基于转录组数据获得了白细胞介素-1 受体相关激酶-b（命名为 McIRAK-b）和白细胞介素-1 受体相关激酶-a（命名为 McIRAK-a），McIRAK-b 的全长为 1815bp，McIRAK-a 为 3168bp，分别编码 532 和 978 个氨基酸。BLASTp 分析和系统发育关系强烈表明，McIRAK-b 的推导氨基酸序列与 IRAK-4 具有高度同源性，而 McIRAK-a 与其他软体动物的 IRAK-1 相似，尤其是在它们的功能域。在包括贻贝、血细胞、鳃、性腺和肝胰腺在内的六种组织中检测到 McIRAK-b 和 McIRAK-a 的表达，在鳃中表达最高。在细菌感染后，McIRAK-b 和 McIRAK-a 在鳃中的表达呈现时间依赖性。在受到溶藻弧菌的刺激后，McIRAK-b 的表达显著增加，在 8 小时时达到峰值（与对照组相比增加了 9.47 倍），而在感染副溶血弧菌时，峰值出现在 4 小时（与对照组相比增加了 12.02 倍）。在受到溶藻弧菌的刺激后，McIRAK-a 的 mRNA 最高点出现在 12 小时（5.16 倍），在受到副溶血弧菌的刺激后，8 小时时达到最高点（4.21 倍）。结果表明，IRAK-b 和 IRAK-a 可能在贻贝的免疫信号通路中发挥重要作用。在 BL21(DE3) 中表达并通过 Ni-NAT Superflow 树脂纯化的 McIRAK-b 和 McIRAK-a 的激酶功能域序列（S_TKc）符合预期的分子量，具有许多活性位点，赋予它们蛋白-蛋白相互作用的功能。本研究可能为 IRAKs 家族在双壳类固有免疫系统中的调控机制提供一些进一步的认识。
