Identification and expression of TRAF6 (TNF receptor-associated factor 6) gene in Zhikong scallop Chlamys farreri.
Identification and expression of TRAF6 (TNF receptor-associated factor 6) gene in Zhikong scallop Chlamys farreri.
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DOI：
10.1016/j.fsi.2008.10.010
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发表时间：
2009-03
影响因子：
4.7
通讯作者：
L. Qiu;L. Song;Yundong Yu;Jianmin Zhao;Lingling Wang;Qing‐Chun Zhang
中科院分区：
文献类型：
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作者：
L. Qiu;L. Song;Yundong Yu;Jianmin Zhao;Lingling Wang;Qing‐Chun Zhang
肿瘤坏死因子受体相关因子6(TRAF6)是TNFR超家族和IL-1R/TLR家族共有的关键信号转导分子，不仅对TNFR超家族的多种生理过程和功能起重要作用，而且还参与获得性免疫和先天免疫。本研究首次在栉孔扇贝中发现了TRAF6基因(命名为CfTRAF6)，并对其进行了鉴定。CfTRAF6的全长为2510bp，包括337bp的5‘端非翻译区，208bp的3’端非翻译区，以及一个编码655个氨基酸的多肽的开放阅读框。预测的CfTRAF6氨基酸序列包含TRAF蛋白的特征基序，包括一个环型锌指、两个TRAF型锌指、一个卷曲区域和一个MATH(meprin和TRAF同源)结构域。CfTRAF6与其他TRAF6的氨基酸序列同源性为28-68%。CfTRAF6序列与其他生物的TRAF序列的系统发育分析表明，CfTRAF6是一个真正的TRAF6同源物。用Real-Time RT-PCR检测CfTRAF6在不同组织中的表达。转录本在血细胞、肌肉、外套膜、心脏、性腺和鳃组织中有结构性表达，但在性腺中表达最高。用20μGML−1和0.5μGML−1肽聚糖(PGN)处理混合原代培养的血细胞，记录CfTRAF6mRNA的表达。0.5μGML−1PGN处理后1.5h~3h，CfTRAF基因的表达水平下降，随后恢复到原来的水平。而CfTRAF6的表达在20μ−1PGN处理后明显降低，并在3h降至最低点(仅为对照的1/9倍)。提示PgN对CfTRAF6有很大的调节作用，可能参与了信号转导和免疫应答。
Tumor necrosis factor receptor-associated factor 6 (TRAF6), a key signaling adaptor molecule common to the TNFR superfamily and IL-1R/TLR family, is important not only for a diverse array of physiological processes functions of the TNFR superfamily, but also is involved in adaptive immunity and innate immunity. In this report, the first bivalve TRAF6 (named as CfTRAF6) gene is identified and characterized from Zhikong scallop Chlamys farreri. The full-length cDNA of CfTRAF6 is of 2510bp, consisting of a 5′-terminal untranslated region (UTR) of 337bp, a 3′-terminal UTR of 208bp with a canonical polyadenylation signal sequence AATAAA and a poly (A) tail, and an open reading frame (ORF) encoding a polypeptide of 655 amino acids. The predicted amino acid sequence of CfTRAF6 comprises characteristic motifs of the TRAF proteins, including a Zinc finger of RING-type, two Zinc fingers of TRAF-type, a coiled-coil region, and a MATH (the meprin and TRAF homology) domain. The overall amino acid sequence identity between CfTRAF6 and other TRAF6s is 28–68%. Phylogenetic analyses of CfTRAF6 sequence with TRAF sequences from other organisms indicate that CfTRAF6 is a true TRAF6 orthologue. The mRNA expression of CfTRAF6 in various tissues is measured by Real-time RT-PCR. The mRNA transcripts are constitutively expressed in tissues of haemocyte, muscle, mantle, heart, gonad and gill, but the highest expression is observed in the gonad. The temporal expressions of CfTRAF6 mRNA in the mixed primary cultured haemocytes are recorded after treatment with 20μgmL−1and 0.5μgmL−1peptidoglycan (PGN). The expression level of CfTRAF mRNA is down-regulated from 1.5h to 3h after the treatment with 0.5μgmL−1PGN, and then recovers to the original level. While the expression of CfTRAF6 is obviously decreased after treatment with 20μgmL−1PGN, and reach the lowest point (only about 1/9 times to control) at 3h. The result suggests that CfTRAF6 can be greatly regulated by PGN and it may be involved in signal transduction and immune response of scallop.