A high mobility group box 1 (HMGB1) gene from Chlamys farreri and the DNA-binding ability and pro-inflammatory activity of its recombinant protein.
A high mobility group box 1 (HMGB1) gene from Chlamys farreri and the DNA-binding ability and pro-inflammatory activity of its recombinant protein.
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DOI：
10.1016/j.fsi.2013.12.008
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发表时间：
2014-02
影响因子：
4.7
通讯作者：
Mengqiang Wang;Lingling Wang;Ying Guo;Zhi Zhou;Qilin Yi;Daoxiang Zhang;Huan Zhang;Rui Liu
中科院分区：
文献类型：
--
作者：
Mengqiang Wang;Lingling Wang;Ying Guo;Zhi Zhou;Qilin Yi;Daoxiang Zhang;Huan Zhang;Rui Liu
高迁移率组框1 （HMGB1）蛋白是一种高度保守的DNA结合蛋白，在维持核小体结构、转录和炎症中起重要作用。本研究利用cDNA末端快速扩增（RACE）技术和表达序列标记（EST）分析，克隆了一段全长1268 bp的直孔扇贝栉孔扇贝（scallopChlamys farreriHMGB1，设计为cfhmgb1） cDNA。cfhmgb1全长cDNA序列包含648 bp的开放阅读框（ORF），编码215个氨基酸的蛋白。cfhmgb1的氨基酸序列与其他已鉴定的hmgb1具有53-57%的相似性。cfhmgb1的氨基酸序列存在2个HMG结构域、2个低复杂度区和1个保守的酸性尾部。cfhmgb1 mRNA转录本在血球、肌肉、套膜、鳃、肝胰脏、肾脏和性腺等组织中均有组成性表达，其中肝胰脏表达量最高。不同病原体相关分子模式（pamp），包括脂多糖（LPS）、肽聚糖（PGN）和葡聚糖（Glu）刺激后，cfhmgb1在血细胞中的mRNA表达谱相似，均在早期上调，然后恢复到原始水平。在扇贝血细胞混合原代培养中，重组cfhmgb1蛋白可结合双链DNA，诱导TNF-α活性释放。综上所述，具有dna结合能力和促炎活性的cfhmgb1可能在扇贝的免疫应答中发挥重要作用。
High-mobility group box 1 (HMGB1) protein, a highly conserved DNA binding protein, plays an important role in maintaining nucleosome structures, transcription, and inflammation. In the present research, a cDNA of 1268 bp for the Zhikong scallopChlamys farreriHMGB1 (designed asCfHMGB1) was cloned via rapid amplification of cDNA ends (RACE) technique and expression sequence tag (EST) analysis. The complete cDNA sequence ofCfHMGB1 contained an open reading frame (ORF) of 648 bp, which encoded a protein of 215 amino acids. The amino acid sequence ofCfHMGB1 shared 53–57% similarity with other identified HMGB1s. There were two HMG domains, two low complexity regions and a conserved acidic tail in the amino acid sequence ofCfHMGB1. The mRNA transcripts ofCfHMGB1 were constitutively expressed in all the tested tissues, including haemocytes, muscle, mantle, gill, hepatopancreas, kidney and gonad, with the highest expression level in hepatopancreas. The mRNA expression profiles ofCfHMGB1 in haemocytes after the stimulation with different pathogen-associated molecular patterns (PAMPs), including lipopolysaccharide (LPS), peptidoglycan (PGN) and glucan (Glu), were similar with an up-regulation in the early stage and then recovered to the original level. The recombinantCfHMGB1 protein could bind double-stranded DNA and induce the release of TNF-α activity in mixed primary culture of scallop haemocytes. These results collectively indicated thatCfHMGB1, with DNA-binding ability and pro-inflammatory activity, could play an important role in the immune response of scallops.