College of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou, 510642, China; Henry Fok School of Biology and Agriculture, Shaoguan University, Shaoguan, 512005, China.
College of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou, 510642, China.
Fish Shellfish Immunol. 2023 Dec;143:109229. doi: 10.1016/j.fsi.2023.109229. Epub 2023 Nov 14.
Rab32 is a member of the Rab GTPase family that is involved in membrane trafficking and immune response, which are crucial for controlling pathogen infection. However, the role of Rab32 in virus infection is not well understood. In this study, we focused on the regulation of Rab32 on virus infection and the host immunity in orange-spotted grouper, Epinephelus coioides. EcRab32 encoded a 213-amino acid polypeptide, which shared a high sequence identity with other Rab32 proteins from fishes to mammals. In healthy orange-spotted grouper, the mRNA of EcRab32 was expressed in all the detected tissues, with the more expression levels in the head kidney, liver and gill. Upon SGIV infection, the expression of EcRab32 was significantly up-regulated in vitro, indicating its potential role in viral infection. EcRab32 was observed to be distributed in the cytoplasm as punctate and vesicle-like structures. EcRab32 overexpression was found to notably inhibit SGIV infection, while the interruption of EcRab32 significantly promoted SGIV infection. In addition, using single particle imaging analysis, we found that EcRab32 overexpression prominently reduced the attachment and internalization of SGIV particles. Furthermore, the results demonstrated that EcRab32 played a positive role in regulating the interferon immune and inflammatory responses. Taken together, these findings indicated that EcRab32 influenced SGIV infection by regulating the host immune response, providing an overall understanding of the interplay between the Rab32 and innate immunity.
Rab32 是 Rab GTPase 家族的成员，参与膜运输和免疫反应，这对于控制病原体感染至关重要。然而，Rab32 在病毒感染中的作用还不是很清楚。在本研究中，我们专注于 Rab32 对感染病毒的橙点石斑鱼（Epinephelus coioides）的宿主免疫反应的调控。EcRab32 编码一个 213 个氨基酸的多肽，与鱼类到哺乳动物的其他 Rab32 蛋白具有高度的序列同一性。在健康的橙点石斑鱼中，EcRab32 的 mRNA 在所有检测到的组织中都有表达，在头肾、肝和鳃中表达水平更高。在 SGIV 感染后，EcRab32 的表达在体外显著上调，表明其在病毒感染中可能具有作用。EcRab32 观察到分布在细胞质中，呈点状和囊泡样结构。EcRab32 的过表达明显抑制了 SGIV 的感染，而 EcRab32 的中断则显著促进了 SGIV 的感染。此外，通过单颗粒成像分析，我们发现 EcRab32 的过表达显著减少了 SGIV 颗粒的附着和内化。此外，结果表明 EcRab32 在调节干扰素免疫和炎症反应中发挥了积极作用。总之，这些发现表明 EcRab32 通过调节宿主免疫反应影响 SGIV 感染，为 Rab32 与先天免疫之间的相互作用提供了全面的了解。
