Molecular cloning and functional analysis of TRAF6 from Yangzhou great white goose Anser anser. (December 2019)

• Record Type: Journal Article
• Authors: Guo, Yaxin; Xu, Ying; Kang, Xilong; Meng, Chuang; Gu, Dan; Zhou, Yingying; Xiong, Dan; Geng, Shizhong; Jiao, Xinan; Pan, Zhiming
• Is Part Of: Developmental and comparative immunology. Volume 101(2019)
• Publication Date: 2019-12
• Subjects: TRAF6 -- Goose -- Cytokines -- TLR ligands -- Salmonella Enteritidis; Immunology -- Periodicals; Developmental immunology -- Periodicals; 616.079
• DOI: 10.1016/j.dci.2019.103435
• Languages: English
• ISSNs: 0145-305X

Abstract

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. Highlights: The structural and functional characteristics of the goTRAF6 gene were conserved and resembled those of other birds. GoTRAF6 mRNA was broadly expressed in all goose tissues examined, especially in heart and muscle. Triggering goTRAF6 led to robust up-regulation of NF-κB activation and IFN-β expression. The expression of goTRAF6 and cytokines were significantly up-regulated in goose PBMCs after TLR ligands stimulation. The mRNA levels of goTRAF6 and cytokines were increased in all examined organs after Salmonella Enteritidis challenge.