LGP2 of black carp plays an important role in the innate immune response against SVCV and GCRV
LGP2 of black carp plays an important role in the innate immune response against SVCV and GCRV
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青鱼的LGP2在针对SVCV和GCRV的先天免疫反应中发挥重要作用
DOI：
10.1016/j.fsi.2016.08.031
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发表时间：
2016
影响因子：
4.7
通讯作者：
Feng Hao
中科院分区：
文献类型：
--
作者：
Xiao Jun;Yan Jun;Chen Hui;Li Jun;Tian Yu;Feng Hao
RIG-I 样受体 (RLR) 检测胞质 RNA 病毒成分，并通过脊椎动物中的下游信号传导启动抗病毒先天免疫反应。本文对青鱼(Mylopharyngodon piceus)的LGP2进行了克隆和鉴定，它是RLR家族的关键成员。青鱼LGP2（bcLGP2）的全长cDNA包含2941个核苷酸，预测的bcLGP2蛋白包含682个氨基酸。 bcLGP2 与 RLR 具有相同的核心同源结构域，包括 N 端 DExD/H 解旋酶结构域、解旋酶超家族 C 端结构域和 C 端调节结构域 (CTD)。在所有选定的组织中均检测到 bcLGP2 mRNA，包括心脏、肝脏、脾脏、肾脏、肠、肌肉、皮肤、鳃；除鳃外，所有组织中 bcLGP2 mRNA 水平均因 GCRV 或 SVCV 感染而增加。青鱼 (MPF) 细胞的 Q-PCR 表明，Poly (I:C) 处理、GCRV 或 SVCV 感染可上调 bcLGP2 转录，但 LPS 或 PMA 处理则不会上调。 Western blot分析表明bcLGP2的分子量约为80 KDa； HeLa细胞和EPC细胞的免疫荧光染色显示bcLGP2是一种胞浆蛋白。转染表达bcLGP2的质粒的EPC细胞对SVCV和GCRV的抗病毒能力明显提高。总的来说，这些数据支持这样的结论：bcLGP2 在宿主抗病毒先天免疫反应中发挥重要作用。
RIG-I like receptors (RLRs) detect cytosolic RNA virus components and initiate antiviral innate immune response through downstream signaling in vertebrates. In this paper, LGP2 of black carp (Mylopharyngodon piceus) has been cloned and characterized, which is a key member of RLR family. The full-length cDNA of black carp LGP2 (bcLGP2) comprises 2941 nucleotides and the predicted bcLGP2 protein contains 682 amino acids. bcLGP2 shares core homologous structural domains of RLRs, including a N-terminnal DExD/H helicase domain, a helicase superfamily c-terminal domain, and a C-terminal regulatory domain (CTD). bcLGP2 mRNA was constitutively detected in all selected tissues including heart, liver, spleen, kidney, intestine, muscle, skin, gill; and bcLGP2 mRNA level was increased in all the tissues except gill in response to GCRV or SVCV infection. Q-PCR ofMylopharyngodon piceusfin (MPF) cells demonstrated that bcLGP2 transcription was up-regulated by Poly (I:C) treatment, GCRV or SVCV infection, but not by LPS or PMA treatment. Western blot analysis demonstrated that the molecular weight of bcLGP2 was around 80 KDa; and the immunofluorescence staining of both HeLa cells and EPC cells showed that bcLGP2 was a cytosolic protein. EPC cells transfected with plasmid expressing bcLGP2 showed obviously improved antiviral ability against SVCV and GCRV. In general, these data support the conclusion that bcLGP2 functions importantly in the host antiviral innate immune response.