Molecular cloning and expression analysis of interferon-inducible transmembrane protein 1 in large yellow croaker Pseudosciaena crocea
Molecular cloning and expression analysis of interferon-inducible transmembrane protein 1 in large yellow croaker Pseudosciaena crocea
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大黄鱼干扰素诱导跨膜蛋白1的分子克隆及表达分析
DOI：
10.1016/j.vetimm.2008.01.004
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发表时间：
2008-07-15
影响因子：
1.8
通讯作者：
Chen, Xinhua
中科院分区：
文献类型：
--
作者：
Wan, Xiang;Chen, Xinhua
在人类细胞中，干扰素诱导的跨膜蛋白1 （IFITM1）是参与同型粘附和抗亲I迭代信号转导的蛋白复合物的一个组成部分。在这里，我们报道了从大黄鱼Pseudosciaena crocea （LycIFITM1）脾脏中克隆的IFITM1同源物。LycIFITM1的完整cDNA全长734个核苷酸（nt），编码124个氨基酸（aa）的蛋白，推测分子量为13.6 kDa。推断的LycIFITM1蛋白与哺乳动物和鱼类中的干扰素诱导跨膜蛋白（IFITMs）具有显著的同源性，具有IFITMs的典型结构特征，包括两个跨膜结构域（分别为43-63和90-112残基）和它们之间的一个胞内结构域（64-89残基），以及一个保守的蛋白激酶C （PKC）磷酸化位点（65-67残基，SIK）。系统发育分析表明，LycIFITM1与鱼类IFITM形成一个簇，反映了与哺乳动物的进化关系相对较远。LycIFITM1基因在鳃、肠、肝、肾、心、脾、肌肉和血液等组织中均有组成性表达。经poly（I:C）诱导后，刺激后24 h， LycIFITM1基因在鳃、肾脏、心脏和脾脏中的表达明显上调，提示LycIFITM1可能参与poly（I:C）诱导的免疫应答。real-time PCR时间过程分析显示，脾和肾中LycIFITM1的mRNA水平受到poly（l:Q）的快速lip调控，并在诱导后24 h达到峰值（脾和肾mRNA分别增加48.7倍和280.4倍）。结果表明，来自大黄鱼的IFITM1同源物可能代表了鱼类中ifitm家族的新成员。(c) 2008 Elsevier B.V.版权所有
In human cells, interferon-inducible transmembrane protein 1 (IFITM1) is a component of protein complexes involved in homotypic adhesion and the transduction of anti pro I iterative signals. Here, we reported the cloning of an IFITM1 homologue from the spleen of large yellow croaker Pseudosciaena crocea (LycIFITM1). The complete cDNA of LycIFITM1 is 734 nucleotides (nt) encoding a protein of 124 amino acids (aa), with a putative molecular weight of 13.6 kDa. The deduced LycIFITM1 protein is significantly homologous to interferon-inducible transmembrane proteins (IFITMs) in mammals and fish, and has the typical structural features of IFITMs, including two transmembrane domains (residues 43-63 and 90-112, respectively) and one intracellular domain between them (residues 64-89), as well as one conserved protein kinase C (PKC) phosphorylation site (residues 65-67, SIK). Phylogenetic analysis showed that LycIFITM1 formed a cluster with fish IFITM, reflecting a relative distant evolutionary relationship from mammals. LycIFITM1 gene was constitutively expressed in various tissues examined, such as gills, intestine, liver, kidney, heart, spleen, muscle and blood. Upon induction with poly(I:C), LycIFITM1 gene expression was obviously up-regulated in gills, kidney, heart and spleen at 24 h after stimulation, suggesting that LycIFITM1 may be involved in the immune response induced by poly(I:C). Time course analysis using real-time PCR showed that the mRNA levels of LycIFITM1 in spleen and kidney were quickly LIP-regulated by poly(l:Q and reached the peak at 24 h post-induction (48.7- and 280.4-fold mRNA increases in spleen and kidney, respectively). The results suggest that the IFITM1 homologue from large yellow croaker may represent a novel member of IFITMs family in fish. (c) 2008 Elsevier B.V. All rights reserved.