Sun Yanan, Wang Jing, Lao Haihua, Yin Zhixin, He Wei, Weng Shaoping, Yu Xiaoqiang, Chan SiuMing, He Jianguo
State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, PR China.
Dev Comp Immunol. 2008;32(4):391-9. doi: 10.1016/j.dci.2007.07.006. Epub 2007 Aug 20.
Apoptosis-associated speck-like protein containing a CARD (ASC) is an adaptor protein that has a bipartite domain structure, an N-terminal PYRIN domain and a C-terminal caspase-recruitment domain (CARD). In this study, we cloned the mandarin fish ASC cDNA (mfASC), which consisted of 899bp with a 115bp 5'-UTR and a 181bp 3'-UTR. The open reading frame encoded 201 amino acids. The mfASC shows 37% identity to an ASC orthologue from zebrafish. The mfASC has two protein-protein interaction domains, an N-terminal PYRIN domain and a C-terminal CARD domain. The mfASC gene structure was determined and had a length of 3954bp with four exons separated by three introns. Northern blot analysis showed that mfASC mRNA is constitutively expressed in the head kidney, gill, hind kidney, spleen and intestine. In vitro studies, mfASC fused with green fluorescent protein appeared as a speck in the transfected 293T cells. When transiently overexpressed in 293T cells, mfASC inhibited NF-kappaB activity with or without tumor necrosis factor (TNFalpha) or lipopolysacharide (LPS) stimulation.
含CARD结构域的凋亡相关斑点样蛋白（ASC）是一种衔接蛋白，具有二分结构域，即N端的PYRIN结构域和C端的半胱天冬酶招募结构域（CARD）。在本研究中，我们克隆了鳜鱼ASC cDNA（mfASC），其长度为899bp，5'-UTR为115bp，3'-UTR为181bp。开放阅读框编码201个氨基酸。mfASC与斑马鱼ASC直系同源物的一致性为37%。mfASC有两个蛋白质-蛋白质相互作用结构域，即N端的PYRIN结构域和C端的CARD结构域。确定了mfASC基因结构，其长度为3954bp，有四个外显子，由三个内含子隔开。Northern印迹分析表明，mfASC mRNA在头肾、鳃、后肾、脾脏和肠道中组成性表达。在体外研究中，与绿色荧光蛋白融合的mfASC在转染的293T细胞中表现为斑点。当在293T细胞中瞬时过表达时，mfASC在有或无肿瘤坏死因子（TNFα）或脂多糖（LPS）刺激的情况下均抑制NF-κB活性。
