Jiangsu Province Engineering Research Center for Aquatic Animals Breeding and Green Efficient Aquacultural Technology, College of Marine Science and Engineering, Nanjing Normal University, 1 Wenyuan Road, Nanjing, Jiangsu, 210023, China.
Jiangsu Province Engineering Research Center for Aquatic Animals Breeding and Green Efficient Aquacultural Technology, College of Marine Science and Engineering, Nanjing Normal University, 1 Wenyuan Road, Nanjing, Jiangsu, 210023, China; Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Sciences, Shandong Normal University, Jinan, Shandong, 250014, China; Co-Innovation Center for Marine Bio-Industry Technology of Jiangsu Province, Lianyungang, Jiangsu, 222005, China.
Mol Immunol. 2020 Oct;126:1-7. doi: 10.1016/j.molimm.2020.07.009. Epub 2020 Jul 23.
Rel/nuclear factor (NF)-κB family of transcription factors paly vital roles in innate immunity response to bacterial and viral infection. Here, we cloned and identified a dorsal homologue (named as MnDorsal) from Macrobrachium nipponense. The full-length cDNA of MnDorsal is 2573 bp with a 1986 bp open reading frame that encodes 661 amino acids. Predicted MnDorsal protein contained a RHD (Rel homology domain), an IPT (Iglike, plexins, and transcriptions factors) domain, and two low complexity regions. Phylogenetic analysis showed that MnDorsal has a closer genetic distance with dorsal homologues from invertebrates. MnDorsal was widely expressed in a variety of tissues, including hemocytes, heart, hepatopancreas, gills, stomach, and intestine. Expression patterns analysis showed that the transcriptional level of MnDorsal in the gills was evidently up-regulated after Staphylococcus aureus, Vibrio parahaemolyticus, white spot syndrome virus, or polyinosinic-polycytidylic acid challenge, suggesting that MnDorsal participates in the immune defenses against pathogens and stimulant challenges. Additionally, the dsRNA-mediated RNA interference analysis showed that knockdown of MnDorsal can significantly inhibit the expression of anti-lipopolysaccharide factor (ALF) and crustin. Further studies revealed that the up-regulated expression of ALFs (MnALF2, MnALF3, and MnALF4) and crustins (MnCrustin3 and MnCrustin4) caused by S. aureus infection were obviously decreased after silencing MnDorsal. These findings suggest that MnDorsal positively regulate the expression of antibacterial peptides (AMPs) during S. aureus infection. Our study will promote to better understand the role of Toll-Dorsal-AMPs pathway in innate immunity response to gram-positive bacterial infection in crustacean.
Rel/nuclear 因子 (NF)-κB 转录因子家族在细菌和病毒感染的固有免疫反应中发挥重要作用。在这里，我们从日本沼虾 (Macrobrachium nipponense) 中克隆并鉴定了一个背侧同源物 (命名为 MnDorsal)。MnDorsal 的全长 cDNA 为 2573bp，具有 1986bp 的开放阅读框，编码 661 个氨基酸。预测的 MnDorsal 蛋白含有一个 RHD（Rel 同源结构域）、一个 IPT（Iglike、plexins 和转录因子）结构域和两个低复杂度区域。系统发育分析表明，MnDorsal 与无脊椎动物的背侧同源物具有更近的遗传距离。MnDorsal 在多种组织中广泛表达，包括血细胞、心脏、肝胰腺、鳃、胃和肠。表达模式分析表明，在金黄色葡萄球菌、副溶血弧菌、白斑综合征病毒或聚肌胞苷酸刺激后，MnDorsal 在鳃中的转录水平明显上调，表明 MnDorsal 参与了对病原体和刺激物的免疫防御。此外，dsRNA 介导的 RNA 干扰分析表明，MnDorsal 的敲低可显著抑制抗脂多糖因子 (ALF) 和几丁质酶的表达。进一步的研究表明，金黄色葡萄球菌感染后，MnDorsal 沉默后，ALFs (MnALF2、MnALF3 和 MnALF4) 和几丁质酶 (MnCrustin3 和 MnCrustin4) 的上调表达明显降低。这些发现表明，MnDorsal 在金黄色葡萄球菌感染期间正向调节抗菌肽 (AMP) 的表达。我们的研究将有助于更好地理解 Toll-Dorsal-AMP 途径在甲壳动物革兰氏阳性菌感染固有免疫反应中的作用。
