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石斑鱼 Atg14 通过抑制宿主固有免疫反应促进新加坡石斑鱼虹彩病毒(SGIV)的复制。

Grouper Atg14 promotes Singapore grouper iridovirus (SGIV) replication by inhibiting the host innate immune response.

机构信息

College of Marine Sciences, South China Agricultural University, Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou, 510642, China; Nansha-South China Agricultural University Fishery Research Institute, Guangzhou, 511400, China.

Nansha-South China Agricultural University Fishery Research Institute, Guangzhou, 511400, China; Department of Biological Sciences, National University of Singapore, Singapore, 117543, Singapore.

出版信息

Fish Shellfish Immunol. 2023 Oct;141:109067. doi: 10.1016/j.fsi.2023.109067. Epub 2023 Sep 7.

Abstract

As one of the important members of the autophagy-related protein family, Atg14 plays a key role in the formation and maturation of autophagosomes. However, little is known about the potential roles of fish Atg14 and its roles in virus infection. In the present study, the homolog of Atg14 (EcAtg14) from the orange-spotted grouper (Epinephelus coioides) was cloned and characterized. The open reading frame (ORF) of EcAtg14 consists of 1530 nucleotides, encoding 509 amino acids, with a predicted molecular weight of 56.9 kDa. EcAtg14 was distributed in all tested tissues, with higher expression in liver, blood and spleen. The expression of EcAtg14 was increased in grouper spleen (GS) cells after Singapore grouper iridovirus (SGIV) infection. EcAtg14 was distributed in the cytoplasm of GS cells. Overexpression of EcAtg14 promoted SGIV replication in GS cells and inhibited IFN3, ISRE and NF-κB promoter activities. Co-immunoprecipitation results showed that there was an interaction between EcAtg14 and EcBeclin. EcAtg14 also promoted the synthesis of LC3-II in GS cells. These findings provide a basis for understanding the innate immune mechanism of grouper against viral infection.

摘要

作为自噬相关蛋白家族的重要成员之一,Atg14 在自噬体的形成和成熟中发挥着关键作用。然而,关于鱼类 Atg14 及其在病毒感染中的作用的潜在作用知之甚少。在本研究中,从橙色斑点石斑鱼(Epinephelus coioides)中克隆并表征了 Atg14 的同源物(EcAtg14)。EcAtg14 的开放阅读框(ORF)由 1530 个核苷酸组成,编码 509 个氨基酸,预测分子量为 56.9 kDa。EcAtg14 分布在所有测试的组织中,在肝脏、血液和脾脏中的表达较高。在 SGIV 感染石斑鱼脾脏(GS)细胞后,EcAtg14 的表达增加。EcAtg14 分布在 GS 细胞的细胞质中。EcAtg14 的过表达促进了 GS 细胞中 SGIV 的复制,并抑制了 IFN3、ISRE 和 NF-κB 启动子活性。免疫共沉淀结果表明 EcAtg14 与 EcBeclin 之间存在相互作用。EcAtg14 还促进了 GS 细胞中 LC3-II 的合成。这些发现为了解石斑鱼对病毒感染的固有免疫机制提供了基础。

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