Zhang Yang, Yu Feng, Li Jun, Tong Ying, Zhang Yuehuan, Yu Ziniu
Key Laboratory of Tropical Marine Bio-resources and Ecology, Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China.
Key Laboratory of Tropical Marine Bio-resources and Ecology, Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China.
Fish Shellfish Immunol. 2014 Oct;40(2):466-71. doi: 10.1016/j.fsi.2014.07.029. Epub 2014 Aug 8.
Retinoic acid-inducible gene I (RIG-I)-like receptor (RLR) is a pivotal receptor that detects numerous RNA and DNA viruses and mediates the innate induction of interferons and pro-inflammatory cytokines upon viral infection. In the present study, we cloned and characterized the first RIG-I gene in a marine mollusk, Crassostrea gigas, and designated it as CgRIG-I. The full-length CgRIG-I cDNA is 3436 bp, including 5'- and 3'-untranslated regions (UTRs) of 93 bp and 286 bp, respectively, and an open reading frame (ORF) of 3057 bp. The gene encodes a 1018 amino acid polypeptide with an estimated molecular mass of 116.5 kDa. SMART analysis showed that the CgRIG-I protein had the typical conserved domains, including the caspase activation and recruitment domains (CARDs), the RNA helicase domain and the C-terminal regulatory domain (RD). Phylogenetic analysis revealed that CgRIG-I was grouped into the clade of its vertebrate homologs. Moreover, CgRIG-I expression could be specifically increased after stimulation by poly(I:C) rather than by other PAMPs such as lipopolysaccharide (LPS), peptidoglycan (PGN), heat-killed Listeria monocytogenes (HKLM) and heat-killed Vibrio alginolyticus (HKVA). Meanwhile, six IRF, three STAT and one NF-κB predicted sites were identified in the CgRIG-I promoter, which was consistent with its high responsiveness to poly(I:C). In summary, this report provides the first CgRIG-I sequence of a mollusk, but its function in the antiviral immune response requires further investigation.
视黄酸诱导基因I(RIG-I)样受体(RLR)是一种关键受体,可检测多种RNA和DNA病毒,并在病毒感染后介导干扰素和促炎细胞因子的先天性诱导。在本研究中,我们克隆并鉴定了海洋软体动物太平洋牡蛎(Crassostrea gigas)中的首个RIG-I基因,并将其命名为CgRIG-I。CgRIG-I cDNA全长3436 bp,分别包括93 bp和286 bp的5'和3'非翻译区(UTR),以及一个3057 bp的开放阅读框(ORF)。该基因编码一个1018个氨基酸的多肽,估计分子量为116.5 kDa。SMART分析表明,CgRIG-I蛋白具有典型的保守结构域,包括半胱天冬酶激活和募集结构域(CARD)、RNA解旋酶结构域和C端调节结构域(RD)。系统发育分析表明,CgRIG-I被归入其脊椎动物同源物的进化枝中。此外,CgRIG-I的表达在受到聚肌胞苷酸(poly(I:C))刺激后可特异性增加,而不是受到其他病原体相关分子模式(PAMP)如脂多糖(LPS)、肽聚糖(PGN)、热灭活的单核细胞增生李斯特菌(HKLM)和热灭活的溶藻弧菌(HKVA)的刺激后增加。同时,在CgRIG-I启动子中鉴定出六个干扰素调节因子(IRF)、三个信号转导子和转录激活子(STAT)以及一个核因子κB(NF-κB)预测位点,这与其对poly(I:C)的高反应性一致。总之,本报告提供了软体动物的首个CgRIG-I序列,但其在抗病毒免疫反应中的功能需要进一步研究。