Abstract：
The Rab proteins primarily regulate vesicular transport between membrane-bound organelles and are important for innate immune. However, there is currently a lack of studies on crustaceans regarding Rab proteins, particularly core Rabs. We identified a Rab11 gene from Procambarus clarkii (PcRab11) and evaluated its potential involvement in immune response. The results showed PcRab11 was 1789 bp long, with an open reading frame of 645 bp encoding 211 amino acids and an estimated molecular weight of 23.8 kDa. Sequence analysis revealed its remarkable evolutionary conservation. The PcRab11 was widely expressed in various tissues, with highest levels in hepatopancreas, and localized within the cell cytoplasm. Upon infection with white spot syndrome virus (WSSV) or Aeromonas veronii, the expression of PcRab11 in immune organs was significantly induced. Furthermore, silencing PcRab11 reduced phagocytosis-related genes expression and haemocytes\' phagocytic activity to FITC-labeled A. veronii, as well as decreased mortality and death time in WSSV or A. veronii infected P. clarkii. Additionally, the potential protein interaction between PcRab11 and 14-3-3ε was identified in haemocytes. Overall, our findings provided evidence for the involvement of Rab11 in P. clarkii\'s immune response, establishing a foundation to explore the immune role of core Rab proteins in crustaceans\' innate immune system.
摘要：
Rab蛋白主要调节膜结合细胞器之间的囊泡转运，对先天免疫很重要。然而,目前缺乏关于甲壳类动物Rab蛋白的研究，尤其是核心Rabs。我们从ClarkiiProcambarusClarkii（PcRab11）中鉴定了Rab11基因，并评估了其在免疫应答中的潜在参与。结果表明，PcRab11长1789bp，具有645bp的开放阅读框，编码211个氨基酸，估计分子量为23.8kDa。序列分析揭示了其显着的进化保守性。PcRab11在各种组织中广泛表达,肝胰腺中含量最高，定位于细胞质内。在感染白斑综合征病毒（WSSV）或维气单胞菌后，显著诱导免疫器官中PcRab11的表达。此外，沉默PcRab11降低了吞噬作用相关基因的表达和血细胞对FITC标记的A.veronii的吞噬活性，以及降低WSSV或A.veronii感染的克氏疟原虫的死亡率和死亡时间。此外,在血细胞中鉴定出PcRab11和14-3-3ε之间的潜在蛋白质相互作用。总的来说,我们的发现为Rab11参与Clarkii的免疫反应提供了证据，为探索核心Rab蛋白在甲壳类动物先天免疫系统中的免疫作用奠定了基础。
