College of Animal Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou, 310058, Zhejiang Province, China.
Dev Comp Immunol. 2020 Dec;113:103788. doi: 10.1016/j.dci.2020.103788. Epub 2020 Jul 18.
Bx42, the homologue of SNW1 in mammals, is involved in pre-mRNA splicing and transcriptional regulation. However, the presence and function of Bx42 have remained poorly understood in invertebrates until now. In the current study, a novel SNW domain-containing protein (MnBx42) from Macrobrachium nipponense was identified, and its potential role in the immune response was investigated. The full-length MnBx42 was 7467 bp with an open reading frame of 1653 bp, encoding 550 amino acids. Real-time PCR analysis suggested that MnBx42 was predominantly expressed in the intestine, gills and hepatopancreas, and immunofluorescence assays indicated that it was located in the nucleus. Its expression level was significantly decreased in M. nipponense post-challenge with white spot syndrome virus (WSSV) as well as Aeromonas hydrophila and Staphylococcus aureus, implying its participation in the innate immune response. The knockdown of MnBx42 in vivo notably increased the susceptibility of the prawns to bacterial infection, markedly increased the bacterial load in the gills, and significantly attenuated the phagocytic activity of haemocytes. Dual-luciferase reporter assays illustrated that MnBx42 could activate the NF-κB pathway. Consistent with this, when MnBx42 was silenced in vivo, the expression levels of antimicrobial peptides (AMPs), including ALF2, ALF3, ALF4, ALF5, Cru1 and Cru2, and NF-κB signalling genes, including dorsal, relish, TAK1, TAB1, Ikkβ, and Ikkε, were significantly reduced. Taken together, these findings may provide new insights about Bx42 in crustaceans and pave the way for a better understanding of the crustacean innate immune system.
Bx42 是哺乳动物中 SNW1 的同源物,参与前体 mRNA 的剪接和转录调控。然而,直到现在,Bx42 在无脊椎动物中的存在和功能仍知之甚少。在本研究中,从日本沼虾中鉴定出一种新型含有 SNW 结构域的蛋白(MnBx42),并研究了其在免疫反应中的潜在作用。MnBx42 的全长为 7467bp,开放阅读框为 1653bp,编码 550 个氨基酸。实时 PCR 分析表明,MnBx42 主要在肠、鳃和肝胰腺中表达,免疫荧光试验表明它位于细胞核中。MnBx42 在日本沼虾感染白斑综合征病毒(WSSV)以及嗜水气单胞菌和金黄色葡萄球菌后表达水平显著降低,表明其参与了先天免疫反应。体内 MnBx42 的敲低显著增加了对细菌感染的易感性,显著增加了鳃中的细菌负荷,并显著降低了血细胞的吞噬活性。双荧光素酶报告基因检测表明,MnBx42 可以激活 NF-κB 通路。与此一致的是,当体内沉默 MnBx42 时,抗菌肽(AMPs),包括 ALF2、ALF3、ALF4、ALF5、Cru1 和 Cru2,以及 NF-κB 信号通路基因,包括 dorsal、relish、TAK1、TAB1、Ikkβ 和 Ikkε 的表达水平显著降低。总之,这些发现可能为甲壳动物中的 Bx42 提供新的见解,并为更好地理解甲壳动物先天免疫系统铺平道路。