Identification of the SAMHD1 gene in grass carp and its roles in inducing apoptosis and inhibiting GCRV proliferation
Identification of the SAMHD1 gene in grass carp and its roles in inducing apoptosis and inhibiting GCRV proliferation
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草鱼SAMHD1基因的鉴定及其诱导细胞凋亡和抑制GCRV增殖的作用
DOI：
10.1016/j.fsi.2019.03.028
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发表时间：
2019-05
影响因子：
4.7
通讯作者：
Hu Chengyu
中科院分区：
文献类型：
--
作者：
Xu Xiaowen;Li Meifeng;Li Dongming;Jiang Zeyin;Liu Changxin;Shi Xiao;Wu Chuxin;Chen Xingxing;Lin Gang;Hu Chengyu
SAMHD 1是一种先天性免疫限制因子，通过IRF 3介导的抗病毒和凋亡反应抑制病毒感染。鱼类SAMHD 1与哺乳动物中的SAMHD 1具有相似的特性。本研究从草鱼（Ctenopharyngodon idellus）中克隆并鉴定了aSAMHD 1的同源基因。CiSAMHD 1的cDNA全长为2792 bp，开放阅读框为1884 bp，编码627个氨基酸。多重序列比对表明SAMHD 1在不同物种间高度保守。系统进化树分析表明，CiSAMHD 1与犀牛金线蛭SAMHD 1具有高度的同源性。表达分析表明，CiSAMHD 1在所检测的脑、眼、脾、鳃、肠、肝、心和肾等组织中广泛表达。经poly I：C处理后，脾脏、肝脏和肠道中的表达显著上调。此外，在CIK细胞中用重组IFN刺激后，可以诱导CiSAMHD 1。鉴定了CiSAMHD 1的启动子序列，以探讨CiSAMHD 1的转录调控机制。CiSAMHD 1的启动子序列（1370 bp）由IRF 1、IRF 3、IRF 9和p65结合元件组成。凝胶迁移率变动分析表明IRF 1、IRF 3、IRF 9和p65蛋白分别与CiSAMHD 1启动子相互作用。双荧光素酶检测和q-PCR结果表明，CiIRF 3和CiIRF 9的过表达能够激活CiSAMHD 1的启动子，而CiIRF 1和Cip 65不能激活CiSAMHD 1的启动子。相反，CiIRF 3或CiIRF 9的敲低抑制CiSAMHD 1的转录。有趣的是，CCK试验表明CiSAMHD 1降低细胞活力。TUNEL凋亡检测和Hoechst 33258染色检测表明，CiSAMHD 1的过表达可诱导细胞凋亡。结晶紫染色、vp 3和vp 5基因检测和病毒滴度测定表明，CiSAMHD 1能抑制草鱼呼肠孤病毒（GCRV）在CIK细胞中的增殖。
SAMHD1 is an innate immunity restriction factor that inhibits virus infection through IRF3-mediated antiviral and apoptotic responses. Fish SAMHD1 shares some similar properties with those in mammals. In this study, aSAMHD1orthologue from grass carp (Ctenopharyngodon idellus) was cloned and characterized. The full-length cDNA ofCiSAMHD1is 2792 bp with an ORF of 1884 bp encoding a polypeptide of 627 amino acids. Multiple alignments showed that SAMHD1 is highly conserved among different species. Phylogenetic tree analysis revealed thatCiSAMHD1shared a high degree of homology withSinocyclocheilus rhinocerous SAMHD1. Expression analysis indicated thatCiSAMHD1was widely expressed in all tissues tested including the brain, eyes, spleen, gill, intestine, liver, heart and kidney. It was significantly up-regulated in spleen, liver and intestines after treatment with poly I:C. Also,CiSAMHD1can be induced following stimulation with recombinant IFN in CIK cells. The promoter sequence ofCiSAMHD1was identified to explore the mechanism underlying the transcriptional regulation ofCiSAMHD1. The promoter sequence ofCiSAMHD1(1370 bp) consists of IRF1, IRF3, IRF9 and p65 binding elements. Gel mobility shift assay also showed that IRF1, IRF3, IRF9 and p65 prokaryotic proteins can separately interact withCiSAMHD1promoter. Dual luciferase assay and q-PCR suggested that the promoter ofCiSAMHD1can be activated by the overexpression ofCiIRF3 andCiIRF9, but cannot be triggered byCiIRF1 andCip65. In contrast, knockdown ofCiIRF3 orCiIRF9 inhibits the transcription ofCiSAMHD1. Intriguingly, CCK assay suggested thatCiSAMHD1 decreased cell viability. TUNEL apoptosis assay and Hoechst 33258 staining assay indicated that apoptosis is induced by the overexpression ofCiSAMHD1. Crystal violet staining, detection of two GCRV genes (vp3 and vp5) and viral titration showed thatCiSAMHD1 can suppress the proliferation of grass carp reovirus (GCRV) in CIK cells.
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影响因子：
4.7
作者：
Q. Lai;Gang Lin;Meisheng Ma;Shenghe Huang;Wen Li;Dongming Li;Meihui Gu;Huiling Mao;Chengyu Hu-C
通讯作者：
Q. Lai;Gang Lin;Meisheng Ma;Shenghe Huang;Wen Li;Dongming Li;Meihui Gu;Huiling Mao;Chengyu Hu-C
影响因子：
82.9
作者：
通讯作者：
--
内质网跨膜蛋白 ZDHHC1 和 STING 均作为硬骨鱼 IRF3 激活的直接接头
影响因子：
4.4
作者：
Xu Xiaowen;Li Meifeng;Wu Zhen;Wang Haizhou;Wang Liqiang;Huang Keyi;Liu Xiancheng;Hou Qunhao;Lin Gang;Hu Chengyu
通讯作者：
Hu Chengyu
DOI：
10.1007/978-3-540-68706-1_1242
发表时间：
2020-02
期刊：
Definitions
影响因子：
--
作者：
D. Hoyer;Eric P Zorrilla;Pietro Cottone;Sarah Parylak;Micaela Morelli;N. Simola;James D. Lane
通讯作者：
D. Hoyer;Eric P Zorrilla;Pietro Cottone;Sarah Parylak;Micaela Morelli;N. Simola;James D. Lane
影响因子：
4.1
作者：
Rao Y;Su J
通讯作者：
Su J