Ding Ding, Chen Xiao-Wei, Kang Li-Hua, Jiang Hai-Shan, Kang Cui-Jie
The Key Laboratory of Plant Cell Engineering and Germplasm Innovation of the Ministry of Education, School of Life Sciences, Shandong University, 27 Shanda South Road, Jinan, Shandong 250100, China; Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, 27 Shanda South Road, Jinan, Shandong 250100, China.
The Key Laboratory of Plant Cell Engineering and Germplasm Innovation of the Ministry of Education, School of Life Sciences, Shandong University, 27 Shanda South Road, Jinan, Shandong 250100, China; Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, 27 Shanda South Road, Jinan, Shandong 250100, China.
Dev Comp Immunol. 2014 Oct;46(2):246-54. doi: 10.1016/j.dci.2014.04.016. Epub 2014 May 2.
The Toll/Toll-like receptor (TLR) signaling pathway has an important role in the innate immunity of animals. Evolutionarily conserved signaling intermediate in Toll pathways (ECSIT) is a protein that functions as an adaptor protein for the Toll/TLR and bone morphogenetic protein signaling pathways. ECSIT is also a key component in the macrophage bactericidal activity of mammals. However, the function of ECSIT in crustaceans remains unclear. In this study, we cloned and identified a functional ECSIT homologue, MjECSIT 1, from kuruma shrimp Marsupenaeus japonicus. The complementary DNA of MjEcsit 1 is 1442 base pairs long, with an open reading frame of 1221 base pairs that encodes a 407-residue polypeptide. Transcripts of MjEcsit 1 are detected in hemocytes, gills, hepatopancreas, stomach, heart, intestines, testes, and ovaries. Such transcripts are upregulated by Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Vibrio anguillarum) injections. The knockdown of MjEcsit 1 by double-stranded RNA injection increases the sensitivity of M. japonicus to S. aureus challenge and weakens the bacterial clearance ability of M. japonicus in vivo. In addition, suppressing MjEcsit 1 restrains the upregulation of two anti-lipopolysaccharide factors by S. aureus injection. The results indicate that MjECSIT 1 is important in the antibacterial immunity of M. japonicus.
Toll/ Toll样受体(TLR)信号通路在动物的先天免疫中起着重要作用。Toll通路中的进化保守信号中间体(ECSIT)是一种蛋白质,作为Toll/TLR和骨形态发生蛋白信号通路的衔接蛋白发挥作用。ECSIT也是哺乳动物巨噬细胞杀菌活性的关键成分。然而,ECSIT在甲壳类动物中的功能仍不清楚。在本研究中,我们从日本对虾(Marsupenaeus japonicus)中克隆并鉴定了一个功能性ECSIT同源物MjECSIT 1。MjEcsit 1的互补DNA长1442个碱基对,开放阅读框为1221个碱基对,编码一个407个氨基酸残基的多肽。MjEcsit 1的转录本在血细胞、鳃、肝胰腺、胃、心脏、肠道、睾丸和卵巢中均有检测到。革兰氏阳性菌(金黄色葡萄球菌)和革兰氏阴性菌(鳗弧菌)注射可使这些转录本上调。通过注射双链RNA敲低MjEcsit 1会增加日本对虾对金黄色葡萄球菌攻击的敏感性,并削弱其体内细菌清除能力。此外,抑制MjEcsit 1会抑制金黄色葡萄球菌注射引起的两种抗脂多糖因子的上调。结果表明,MjECSIT 1在日本对虾的抗菌免疫中起重要作用。