Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China.
Sci Rep. 2020 Sep 17;10(1):15315. doi: 10.1038/s41598-020-72311-0.
Duck reovirus (DRV) is a fatal member of the genus Orthoreovirus in the family Reoviridae. The disease caused by DRV leads to huge economic losses to the duck industry. Post-translational modification is an efficient strategy to enhance the immune responses to virus infection. However, the roles of protein phosphorylation in the responses of ducklings to Classic/Novel DRV (C/NDRV) infections are largely unknown. Using a high-resolution LC-MS/MS integrated to highly sensitive immune-affinity antibody method, phosphoproteomes of Cairna moschata spleen tissues under the C/NDRV infections were analyzed, producing a total of 8,504 phosphorylation sites on 2,853 proteins. After normalization with proteomic data, 392 sites on 288 proteins and 484 sites on 342 proteins were significantly changed under the C/NDRV infections, respectively. To characterize the differentially phosphorylated proteins (DPPs), a systematic bioinformatics analyses including Gene Ontology annotation, domain annotation, subcellular localization, and Kyoto Encyclopedia of Genes and Genomes pathway annotation were performed. Two important serine protease system-related proteins, coagulation factor X and fibrinogen α-chain, were identified as phosphorylated proteins, suggesting an involvement of blood coagulation under the C/NDRV infections. Furthermore, 16 proteins involving the intracellular signaling pathways of pattern-recognition receptors were identified as phosphorylated proteins. Changes in the phosphorylation levels of MyD88, NF-κB, RIP1, MDA5 and IRF7 suggested a crucial role of protein phosphorylation in host immune responses of C. moschata. Our study provides new insights into the responses of ducklings to the C/NDRV infections at PTM level.
鸭呼肠孤病毒（DRV）是呼肠孤病毒科呼肠孤病毒属的一种致命成员。由 DRV 引起的疾病给养鸭业造成了巨大的经济损失。翻译后修饰是增强病毒感染免疫反应的有效策略。然而，蛋白质磷酸化在雏鸭对经典/新型 DRV（C/NDRV）感染反应中的作用在很大程度上尚不清楚。本研究使用高分辨率 LC-MS/MS 与高灵敏度免疫亲和抗体法相结合，分析了 C/NDRV 感染下番鸭脾脏组织的磷酸蛋白质组，共鉴定到 2853 个蛋白上的 8504 个磷酸化位点。在与蛋白质组数据归一化后，C/NDRV 感染下分别有 288 个蛋白上的 392 个和 342 个蛋白上的 484 个磷酸化位点发生显著变化。为了表征差异磷酸化蛋白（DPP），进行了系统的生物信息学分析，包括基因本体注释、结构域注释、亚细胞定位和京都基因与基因组百科全书途径注释。鉴定到两个重要的丝氨酸蛋白酶系统相关蛋白，凝血因子 X 和纤维蛋白原α链，作为磷酸化蛋白，表明凝血在 C/NDRV 感染下可能参与其中。此外，还鉴定到 16 个涉及模式识别受体的细胞内信号通路的蛋白作为磷酸化蛋白。MyD88、NF-κB、RIP1、MDA5 和 IRF7 的磷酸化水平变化表明蛋白质磷酸化在宿主对 C/NDRV 的免疫反应中起着至关重要的作用。本研究在翻译后修饰水平上为鸭对 C/NDRV 感染的反应提供了新的见解。
