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扬州鹅 Anser anser 中 TRAF6 的分子克隆与功能分析

Molecular cloning and functional analysis of TRAF6 from Yangzhou great white goose Anser anser.

机构信息

Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Jiangsu, 225009, China; Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, Jiangsu, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Jiangsu, China.

Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Jiangsu, 225009, China; Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, Jiangsu, 225009, China; Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, Jiangsu, China; Joint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Jiangsu, China.

出版信息

Dev Comp Immunol. 2019 Dec;101:103435. doi: 10.1016/j.dci.2019.103435. Epub 2019 Jul 6.

Abstract

TNF receptor-associated factor 6 (TRAF6) is an adaptor protein and an E3 ubiquitin ligase mediating multiple cell signaling pathway activation in a context-dependent manner. TRAF6 plays critical roles in innate immune response and regulates function of antigen-presenting cells. Here, we cloned the goose TRAF6 (goTRAF6) gene from a healthy Yangzhou great white goose (Anser anser), which had a typical TRAF structure and shared a high-sequence identity with TRAF6 of other birds. Quantitative real-time PCR revealed that goTRAF6 mRNA was broadly expressed in all the studied tissues, with highest expression in the heart and pectoral muscle. Overexpression of goTRAF6 caused NF-κB activation in a dose-dependent manner and substantially upregulated IFN-β expression in HEK293T cells. Following Toll-like receptor (TLR) ligand stimulation of goose peripheral blood mononuclear cells, goTRAF6 and downstream inflammatory cytokine mRNA levels considerably up-regulated, especially at early stages. Salmonella Enteritidis challenge caused overexpression of goTRAF6 and cytokine mRNA in all the examined organs. These findings demonstrated that goTRAF6 played a substantial role in TLR-TRAF6 signaling cascade, and further contributed to the antibacterial-responses in host.

摘要

肿瘤坏死因子受体相关因子 6(TRAF6)是一种衔接蛋白和 E3 泛素连接酶,以依赖于上下文的方式介导多种细胞信号通路的激活。TRAF6 在先天免疫反应中发挥关键作用,并调节抗原呈递细胞的功能。在这里,我们从健康扬州鹅(Anser anser)中克隆了鹅 TRAF6(goTRAF6)基因,其具有典型的 TRAF 结构,并与其他鸟类的 TRAF6 具有高度的序列同一性。实时定量 PCR 显示,goTRAF6 mRNA 在所有研究的组织中广泛表达,在心脏和胸肌中表达最高。goTRAF6 的过表达以剂量依赖性方式引起 NF-κB 激活,并在 HEK293T 细胞中显著上调 IFN-β 的表达。在鹅外周血单核细胞受到 Toll 样受体(TLR)配体刺激后,goTRAF6 和下游炎症细胞因子 mRNA 水平显著上调,尤其是在早期阶段。肠炎沙门氏菌攻毒导致所有检测器官中 goTRAF6 和细胞因子 mRNA 的过度表达。这些发现表明,goTRAF6 在 TLR-TRAF6 信号级联中起重要作用,并进一步有助于宿主的抗菌反应。

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