Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture and Rural Affairs, Fisheries College, Jimei University, Xiamen 361021, China.
Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China.
Int J Mol Sci. 2022 Jun 9;23(12):6475. doi: 10.3390/ijms23126475.
Phosphoribosyl pyrophosphate synthetases (EC 2.7.6.1) are key enzymes in the biological synthesis of phosphoribosyl pyrophosphate and are involved in diverse developmental processes. In our previous study, the gene was discovered as a key disease-resistance candidate gene in yellow drum, , in response to the infection of , through genome-wide association analysis. This study mainly focused on the characteristics and its roles in immune responses of the gene in yellow drum. In the present study, the gene was cloned from yellow drum, encoding a protein of 320 amino acids. Bioinformatic analysis showed that PRPS1 was highly conserved during evolution. Quantitative RT-PCR demonstrated that was highly expressed in the head-kidney and brain, and its transcription and translation were significantly activated by infection examined by RT-qPCR and immunohistochemistry analysis, respectively. Subcellular localization revealed that PRPS1 was localized in cytoplasm. In addition, semi-in vivo pull-down assay coupled with mass spectrometry identified myeloid differentiation factor 88 (MyD88) as an PRPS1-interacting patterner, and their interaction was further supported by reciprocal pull-down assay and co-immunoprecipitation. The inducible expression of by suggested that the linker molecule MyD88 in innate immune response may play together with PRPS1 to coordinate the immune signaling in yellow drum in response to the pathogenic infection. We provide new insights into important functions of PRPS1, especially PRPS1 in the innate immunity of teleost fishes, which will benefit the development of marine fish aquaculture.
磷酸核糖焦磷酸合成酶（EC 2.7.6.1）是生物合成磷酸核糖焦磷酸的关键酶，参与多种发育过程。在我们之前的研究中，通过全基因组关联分析，发现 基因是大黄鱼对 感染产生抗性的关键候选基因。本研究主要集中于大黄鱼 基因的特征及其在免疫反应中的作用。本研究从大黄鱼中克隆出 基因，编码一个由 320 个氨基酸组成的蛋白质。生物信息学分析表明，PRPS1 在进化过程中高度保守。定量 RT-PCR 显示，在头部-肾脏和大脑中高度表达，通过 RT-qPCR 和免疫组织化学分析分别检测到其转录和翻译在 感染后显著激活。亚细胞定位显示 PRPS1 定位于细胞质。此外，半体内下拉实验结合质谱鉴定髓样分化因子 88（MyD88）为 PRPS1 的相互作用模式识别物，通过相互下拉实验和共免疫沉淀进一步证实了它们的相互作用。 的诱导表达表明，先天免疫反应中的衔接分子 MyD88 可能与 PRPS1 一起协调大黄鱼对病原感染的免疫信号。我们为 PRPS1 的重要功能提供了新的见解，特别是 PRPS1 在硬骨鱼类先天免疫中的作用，这将有助于海洋鱼类养殖的发展。
