Abstract：
The tumor necrosis factor (TNF) receptor-associated factor (TRAF) family has been reported to be involved in many immune pathways. In a previous study, we identified 5 TRAF genes, including TRAF2, 3, 4, 6, and 7, in the bay scallop (Argopecten irradians, Air) and the Peruvian scallop (Argopecten purpuratus, Apu). Since TRAF6 is a key molecular link in the TNF superfamily, we conducted a series of studies targeting the TRAF6 gene in the Air and Apu scallops as well as their hybrid progeny, Aip (Air ♀ × Apu ♂) and Api (Apu ♀ × Air ♂). Subcellular localization assay showed that the Air-, Aip-, and Api-TRAF6 were widely distributed in the cytoplasm of the human embryonic kidney cell line (HEK293T). Additionally, dual-luciferase reporter assay revealed that among TRAF3, TRAF4, and TRAF6, only the overexpression of TRAF6 significantly activated NF-κB activity in the HEK293T cells in a dose-dependent manner. These results suggest a crucial role of TRAF6 in the immune response in Argopecten scallops. To investigate the specific immune mechanism of TRAF6 in Argopecten scallops, we conducted TRAF6 knockdown using RNA interference. Transcriptomic analyses of the TRAF6 RNAi and control groups identified 1194, 2403, and 1099 differentially expressed genes (DEGs) in the Air, Aip, and Api scallops, respectively. KEGG enrichment analyses revealed that these DEGs were primarily enriched in transport and catabolism, amino acid metabolism, peroxisome, lysosome, and phagosome pathways. Expression profiles of 28 key DEGs were confirmed by qRT-PCR assays. The results of this study may provide insights into the immune mechanisms of TRAF in Argopecten scallops and ultimately benefit scallop breeding.
摘要：
据报道，肿瘤坏死因子（TNF）受体相关因子（TRAF）家族参与许多免疫途径。在之前的研究中,我们鉴定了5个TRAF基因,包括TRAF2、3、4、6和7，在海湾扇贝(Argopectenirrangans，空气）和秘鲁扇贝（紫癜，阿普）。由于TRAF6是TNF超家族中的关键分子链接，我们在Air和Apu扇贝及其杂种后代中进行了一系列针对TRAF6基因的研究，Aip(Air‰×Apu‰)和Api(Apu‰×Air‰)。亚细胞定位实验表明，空气-，AIP-,Api-TRAF6广泛分布在人胚肾细胞系(HEK293T)的细胞质中。此外,双荧光素酶报告基因分析显示，在TRAF3，TRAF4和TRAF6中，只有TRAF6的过表达以剂量依赖性方式显着激活HEK293T细胞中的NF-κB活性。这些结果表明，TRAF6在卵裂扇贝的免疫反应中起着至关重要的作用。探讨TRAF6在扇贝中的特异性免疫机制，我们使用RNA干扰进行TRAF6敲低。TRAF6RNAi和对照组的转录组学分析鉴定了空气中的1194、2403和1099差异表达基因（DEGs），Aip,和Api扇贝，分别。KEGG富集分析表明，这些DEG主要富集在运输和分解代谢方面，氨基酸代谢，过氧化物酶体,溶酶体,和吞噬路径。通过qRT-PCR测定确认28个关键DEGs的表达谱。这项研究的结果可能提供对扇贝中TRAF的免疫机制的见解，并最终有利于扇贝的育种。
