Molecular characterization, expression of chicken TBK1 gene and its effect on IRF3 signaling pathwayReport as inadecuate

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TRAF family member-associated NF-κB activator TANK-binding kinase1 TBK1 is a serine-threonine kinase at the crossroads of multiple interferon IFN-inducing signaling pathways in innate immunity. The importance of TBK1 in antiviral immunity is well established in mammal models, but in chicken, the molecular characterization and potential function of TBK1 remain unclear. In the present study, the open-reading frame ORF of chicken TBK1 chTBK1 was cloned and characterized. The sequencing results revealed that the chTBK1 ORF consists of 2190 base pairs bp encoding a deduced protein of 729 amino acid residues. Multiple sequence alignment analysis demonstrated chTBK1 similarity to other birds and mammals, which indicates that it is evolutionarily conserved. Quantitative real-time PCR qRT-PCR results showed that chTBK1 was ubiquitously expressed in chicken tissues and expression was especially high in immune tissues. In addition, the expression of chTBK1 was significantly up-regulated by infection with avian leukosis virus subgroup J ALV-J both in vivo and in chicken embryo fibroblasts CEFs challenged with ALV-J or stimulated with poly I:C in vitro. Consistent with the activation of chTBK1, the interferon regulatory factor 3 IRF3 and IFNβ gene in CEFs were also up-regulated after challenge with ALV-J or polyI:C. In contrast, the expression of IRF3 and IFNβ in CEFs was significantly reduced by siRNA targeting the chTBK1 gene compared with a negative control NC during ALV-J infection or polyI:C transfection. In conclusion, our results demonstrated that chTBK1 may be an important immunoregulator for IRF3 and IFNβ induction in response to viral stimulation in chicken.

Author: Yan Wang , Yue Yin , Xi Lan , Fei Ye , Kai Tian, Xiaoling Zhao, Huadong Yin, Diyan Li, Hengyong Xu, Yiping Liu, Qing Zhu



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