Identification and characterization of two homologues of interferon-stimulated gene ISG15 in crucian carp
Identification and characterization of two homologues of interferon-stimulated gene ISG15 in crucian carp
复制标题
鲫鱼干扰素刺激基因ISG15的两个同源物的鉴定和表征
DOI：
10.1016/j.fsi.2006.09.004
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发表时间：
2007-07-01
影响因子：
4.7
通讯作者：
Gui, Jian-Fang
中科院分区：
文献类型：
--
作者：
Zhang, Yi-Bing;Wang, Ying-Lu;Gui, Jian-Fang
ISG15是病毒感染、干扰素刺激和脂多糖刺激诱导作用最强的基因之一，到目前为止在哺乳动物中只发现了一个拷贝。利用抑制性消减杂交技术，从紫外线灭活的草鱼出血症病毒(GCHV)感染和干扰素产生的CAB细胞(鲫鱼胚胎细胞)中克隆了两个鱼类ISG15基因，命名为CaISG15-1和CaISG15-2。两条鲫鱼ISG15的全长cDNA序列分别编码一个155个氨基酸的蛋白质和一个161个氨基酸的蛋白质，两者具有哺乳动物ISG15蛋白的特征结构，包括两个串联的泛素样结构域和C端典型的LRLRGG基序，两者的同源性为78.9%。在激活病毒、紫外线灭活的GCHV和含干扰素的上清液(ICS)处理的CAB细胞中，CaISG15-1和CaISG15-2的表达均上调，但表现出不同的动力学特征。Poly I：C和脂多糖也能诱导这两个基因的mRNA表达增加。在对激活的GCHV、紫外线灭活的GCHV、CAB ICS、Poly 1：12和内毒素反应的CAB细胞中，CaISG15-1的表达比CaISG15-2的上调更显著。这些结果表明，鲫鱼体内存在两种ISG15同源物，它们在抵抗病毒和细菌感染的天然免疫中可能发挥着不同的作用。(C)2006爱思唯尔有限公司。保留所有权利。
ISG15 is one of the most strongly induced genes upon viral infection, interferon (IFN) stimulation, and lipopolysaccharide, (LPS) stimulation, and only one copy has been found in mammals so far. Here two fish ISG15 genes, termed CaISG15-1 and CaISG15-2, have been cloned and sequenced from UV-inactivated GCHV (grass carp haemorrhagic virus)-infected and IFN-produced CAB cells (crucian carp Carassius auratus blastulae embryonic cells) by suppression subtractive hybridization. The full-length cDNA sequences of two crucian carp ISG15 encode a 155-amino-acid protein and a 161-amino-acid protein, both of which show 78.9% identity overall and possess the characteristic structures of mammalian ISG15 proteins including two tandem ubiquitin-like domains and the C-terminal canonical LRLRGG motif. In CAB cells treated with different stimuli including active virus, UV-inactivated GCHV and IFN containing supernatant (ICS), the expression of both CaISG15-1 and CaISG15-2 was up-regulated but displayed different kinetics. Poly I:C and LPS were also able to induce an increase in mRNA for both genes. In CAB cells responsive to active GCHV, UV-inactivated GCHV, CAB ICS, Poly 1:12 and LPS, CaISG15-1 was upregulated more significantly than CaISG15-2. These results suggest that there are two ISG15 homologues in crucian carp, both of which might play distinct roles in innate immunity against viral and bacterial infection. (c) 2006 Elsevier Ltd. All rights reserved.