PRRSV利用Rab18调控宿主脂质代谢促进病毒增殖的分子机制

为了研究PRRSV与宿主胞内运输系统之间的相互作用，我们前期构建了Rab基因的shRNA文库，筛选参与PRRSV增殖的关键Rab蛋白。其中，脂滴代谢相关Rab被显著富集。发现PRRSV感染显著上调Rab18的表达，促进围脂滴蛋白ADRP的降解，增加胞内游离脂肪酸的含量。为进一步解析Rab18参与PRRSV增殖的机制，本项目拟利用生化与分子生物学方法，解析PRRSV编码蛋白促进Rab18表达的分子机制；其次利用脂滴纯化和抑制剂等方法鉴定Rab18表达量的增加促进ADRP降解的机制，并检测Rab18与ADRP的相互作用；最后利用HPLC和CRISPR/Cas9等方法确定失去ADRP保护的脂滴降解途径，以期阐明PRRSV通过上调Rab18，加快ADRP降解，增加脂解从而促进病毒增殖的机制。这些研究将揭示宿主胞内运输系统及脂质代谢在PRRSV生活周期中的功能及机制，为PRRS的防控提供理论支撑。

To investigate the interaction between PRRSV and the host intracellular transport system, we previously constructed a shRNA library of Rab genes and screened the key Rab proteins involved in PRRSV proliferation. Among them, lipid droplet metabolism related Rabs have been significantly enriched. PRRSV infection up-regulates Rab18 expression, promotes the degradation of lipid droplet coating protein ADRP, and increases the content of intracellular free fatty acids. To accurately analyze the mechanism of Rab18 involved in PRRSV proliferation, we first analyze the molecular mechanism of which PRRSV-encoded protein promotes Rab18 expression by biochemical and molecular biological methods; Secondly, the mechanism of increasing Rab18 expression promoting ADRP degradation will be identified by lipid droplet purification and inhibitors, and the interaction between Rab18 and ADRP will be detected. Finally, lipid droplet degradation pathways will be determined by HPLC and CRISPR/Cas9. This project aims to elucidate the mechanism of PRRSV promoting virus proliferation by up-regulating Rab18, accelerating ADRP degradation and increasing lipolysis. These studies will not only reveal the function and mechanism of host intracellular transport and lipid metabolism in the PRRSV life cycle, but also provide theoretical support for the prevention and control of PRRS.