Key Laboratory of Mariculture & Stock Enhancement in North China's Sea, Ministry of Agriculture, Dalian Ocean University, Dalian, Liaoning 116023, PR China.
Key Laboratory of Mariculture & Stock Enhancement in North China's Sea, Ministry of Agriculture, Dalian Ocean University, Dalian, Liaoning 116023, PR China.
Gene. 2018 Oct 30;675:110-118. doi: 10.1016/j.gene.2018.06.072. Epub 2018 Jun 27.
The nucleotide-binding oligomerization domain-like receptors (NOD-like receptors, NLRs) can regulate the innate immune process and is an important part of inflammatory body. In this study, we use transcriptome sequencing and the rapid amplification of cDNA ends approach to identify a novel NLRP gene in Apostichopus japonicus. We designated the gene as AjNLRP10. The full-length of AjNLRP10 is 4509 bp. The putative open reading frame comprising 3489 bp encodes a polypeptide with 1162 amino acid residues. The predicted molecular mass of AjNLRP10 is 132.87 kDa and its theoretical pI is 5.60. AjNLRP10 comprises a signal peptide with two Ig superfamily (IgSF) domains and a NACHT [NAIP (neuronal apoptosis inhibitory protein), CIITA (MHC class II transcription activator), HET-E (incompatibility locus protein from Podospora anserina) and TP1 (telomerase-associated protein)] domain. Spatial distribution expression analysis detected AjNLRP10 in all of the tissues tested, but with higher expression in the coelomocytes, medium expression in the intestine and respiratory tree, and slightly weaker expression in the body wall, tube feet, and longitudinal muscle. The expression levels of AjNLRP10 in the respiratory tree and intestines of sea cucumbers with skin ulceration syndrome were increased by 4-fold and 2.7-fold compared with those in healthy sea cucumbers, respectively. We investigated expression profiles of AjCasepase-1 (Cysteinyl aspartate specific proteinase-1) and AjMMP37 (mitochondrial protein-37) after AjNLRP10 knock-down and discovered that AjCasepase-1 was raised by 2.60-fold and AjMMP37 was raised by 3.84-fold. The study showed that AjNLRP10 has inhibitory effect in the immune process. In conclusion, this study showed that the AjNLRP10 protein found in the sea cucumber involved with the innate immune responses against bacterial infection. It has a similar structure and biological function to that in other organisms, where it appears to be involved with these results provide insights into the innate immune mechanism in the sea cucumber as well as suggesting new strategies for disease prevention, molecular therapy, and the development of novel drugs for sea cucumbers.
核苷酸结合寡聚化结构域样受体（NOD-like receptors，NLRs）可以调节先天免疫过程，是炎症体的重要组成部分。在这项研究中，我们使用转录组测序和快速扩增 cDNA 末端的方法在刺参中鉴定了一个新的 NLRP 基因。我们将该基因命名为 AjNLRP10。AjNLRP10 的全长为 4509bp。包含 3489bp 的推定开放阅读框编码具有 1162 个氨基酸残基的多肽。AjNLRP10 的预测分子质量为 132.87kDa，理论等电点为 5.60。AjNLRP10 包含一个带有两个免疫球蛋白超家族（IgSF）结构域和一个 NACHT[NAIP（神经元凋亡抑制蛋白）、CIITA（MHC 类 II 转录激活因子）、HET-E（来自 Podospora anserina 的不兼容基因座蛋白）和 TP1（端粒酶相关蛋白）]结构域的信号肽。空间分布表达分析检测到 AjNLRP10 在所有测试的组织中均有表达，但在体腔细胞中的表达较高，在肠和呼吸树中的表达中等，在体壁、管足和纵肌中的表达略弱。与健康海参相比，患有皮肤溃疡综合征的海参呼吸树和肠道中 AjNLRP10 的表达水平分别增加了 4 倍和 2.7 倍。在 AjNLRP10 敲低后，我们研究了 AjCasepase-1（半胱天冬氨酸特异性蛋白酶-1）和 AjMMP37（线粒体蛋白-37）的表达谱，发现 AjCasepase-1 升高了 2.60 倍，AjMMP37 升高了 3.84 倍。研究表明，AjNLRP10 在免疫过程中具有抑制作用。综上所述，本研究表明，在海参中发现的 AjNLRP10 蛋白参与了对细菌感染的先天免疫反应。它具有与其他生物相似的结构和生物学功能，似乎与这些结果有关，为海参的先天免疫机制提供了新的见解，并为海参的疾病预防、分子治疗和新型药物的开发提供了新的策略。
