Cloning and characterization of a LPS-regulatory gene having an LPS binding domain in kuruma prawn Marsupenaeus japonicus
Cloning and characterization of a LPS-regulatory gene having an LPS binding domain in kuruma prawn Marsupenaeus japonicus
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DOI：
10.1016/j.molimm.2005.12.009
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发表时间：
2006-05-01
影响因子：
3.6
通讯作者：
Soma, Gen-Ichiro
中科院分区：
文献类型：
--
作者：
Nagoshi, Hiroki;Inagawa, Hiroyuki;Soma, Gen-Ichiro
LPS 被认为是多种动物免疫系统的有效刺激剂，包括哺乳动物和鲎 (HSC)。这两个动物群体都具有 LPS 反应的抑制性调节蛋白，例如哺乳动物中的杀菌/通透性增加蛋白和 HSC 中的抗 LPS 因子 (ALF)。对虾是一种有价值的水产养殖物种，但对 LPS 作出反应的调节分子和/或机制在很大程度上尚不清楚。为了研究库鲁玛虾 LPS 反应的分子机制，我们克隆了具有 LPS 结合域的 cDNA。全长 cDNA 基因。克隆了一条长746 bp、编码123个氨基酸残基的M-ALF（Marsupenaeus japonicus ALF样肽）。该基因的 3' 非翻译区有 ATTTA 五聚体重复四次：这被称为信使 RNA 稳定序列。推导的氨基酸序列与日本HSC-ALF有42%的同源性。特别是，两者都具有碱性和疏水性氨基酸簇，表明该区域可能与脂质 A 结合。通过 RT-PCR 测定了血细胞、淋巴器官、心脏、肠和鳃的 mRNA 表达。给予LPS后1.5-3小时，淋巴器官中mRNA表达增加，但在6小时后降至正常水平。含有Cys30至Cys51的合成肽对RAW264.7细胞中的鲎反应和NO产生具有LPS中和活性。这些数据表明，在克鲁玛虾中，M-ALF 在病原体入侵后的急性期反应中充当 LPS 调节剂。 (c) 2005 Elsevier Ltd. 保留所有权利。
LPS is known as an effective stimulator of the immune system in various animals, including mammals and horseshoe crabs (HSC). Both of these animal groups have suppressive regulatory proteins for the LPS response, e.g. the bactericidal/permeability increasing protein in mammals and anti-LPS factor (ALF) in HSC. Prawns are a valuable aquaculture species, but the regulatory molecules and/or mechanisms that respond to LPS are largely unknown. To investigate the molecular mechanism of the LPS response in kuruma prawns, we cloned a cDNA having a LPS binding domain. A full-length cDNA gene. denoted as M-ALF (Marsupenaeus japonicus ALF-like peptide) was cloned that consisted of 746 bp and encoded 123 amino-acid residues. The 3' non-translated region of this gene had the pentamer of ATTTA repeated four times: this is known as sequences for messenger RNA stabilization. Deduced amino-acid sequences showed a 42% homology with Japanese HSC-ALF. In particular, both have clusters of basic and hydrophobic amino acids, indicating that the region is probably binding to lipid A. The mRNA expression was determined for hemocytes, lymphoid organs, hearts, intestines and gills by RT-PCR. The mRNA expression was augmented 1.5-3 h after LPS administration in lymphoid organs, but then decreased to normal level at 6 h. Synthetic peptides containing Cys30 to Cys51 had LPS neutralizing activity to the Limulus reaction and NO production in RAW264.7 cells. These data suggest that in kuruma prawns, M-ALF acts as a LPS regulator during the acute phase response after invasion of pathogens. (c) 2005 Elsevier Ltd. All rights reserved.