Li Yinan, Sun Jiejie, Yang Wenwen, Liu Yu, Lian Xingye, Wang Lingling, Song Linsheng
College of Life Sciences, Liaoning Normal University, Dalian 116029, Liaoning, China; Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian 116023, China; Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian 116023, China; Liaoning Key Laboratory of Aquatic Disease Prevention and Control, Dalin Ocean University, Dalian 116023, China.
Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian 116023, China; Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian 116023, China; Liaoning Key Laboratory of Aquatic Disease Prevention and Control, Dalin Ocean University, Dalian 116023, China.
Int J Biol Macromol. 2025 Apr;302:140470. doi: 10.1016/j.ijbiomac.2025.140470. Epub 2025 Jan 28.
DM9 domain-containing protein (DM9CP) as pattern recognition molecule is involved in regulating the inflammation-related signaling pathway in invertebrates. In the present study, a DM9CP with two tandem DM9 repeats (designated as CgDM9CP-8) was identified from Crassostrea gigas. The mRNA transcript of CgDM9CP-8 was the highest in haemocytes. Recombinant CgDM9CP-8 was able to bind various PAMPs (MAN, PGN and LPS) and microbes (Vibrio splendidus, Staphylococcus aureus, Bacillus subtilis, Pichia pastoris, and Yarrowia lipolytica). CgDM9CP-8 protein was distributed in haemocytes and cell-free haemolymph. After lipopolysaccharide (LPS) or V. splendidus stimulation, the mRNA expressions of CgDM9CP-8 in haemocytes were significantly up-regulated at 3 and 6 h, respectively. rCgDM9CP-8 exhibited binding activity to V. splendidus and E. coli, inhibiting their growth in vitro. CgDM9CP-8 was able to bind CgMASPL-1 both in vivo and in vitro to lead to complement CgC3 cleavage. In addition, in CgDM9CP-8-RNAi oysters, the mRNA expressions of interleukin 17s (CgIL17s) in haemocytes decreased significantly after V. splendidus stimulation. These results suggested that CgDM9CP-8 functioned as a pattern recognition and antibacterial molecule to regulate the mRNA expressions of CgIL17s through the activation of CgC3 in C. gigas.
含DM9结构域蛋白（DM9CP）作为模式识别分子参与调节无脊椎动物炎症相关信号通路。在本研究中，从太平洋牡蛎中鉴定出一种具有两个串联DM9重复序列的DM9CP（命名为CgDM9CP - 8）。CgDM9CP - 8的mRNA转录本在血细胞中最高。重组CgDM9CP - 8能够结合多种病原体相关分子模式（甘露聚糖、肽聚糖和脂多糖）以及微生物（灿烂弧菌、金黄色葡萄球菌、枯草芽孢杆菌、毕赤酵母和解脂耶氏酵母）。CgDM9CP - 8蛋白分布于血细胞和无细胞血淋巴中。脂多糖（LPS）或灿烂弧菌刺激后，血细胞中CgDM9CP - 8的mRNA表达分别在3小时和6小时显著上调。重组CgDM9CP - 8对灿烂弧菌和大肠杆菌具有结合活性，可抑制它们在体外的生长。CgDM9CP - 8在体内和体外均能与CgMASPL - 1结合，导致补体CgC3裂解。此外，在CgDM9CP - 8 - RNAi牡蛎中，灿烂弧菌刺激后血细胞中白细胞介素17（CgIL17s）的mRNA表达显著下降。这些结果表明，CgDM9CP - 8作为模式识别和抗菌分子，通过激活太平洋牡蛎中的CgC3来调节CgIL17s的mRNA表达。
