Two Rab GTPases, EsRab-1 and EsRab-3, involved in anti-bacterial response of Chinese mitten crab Eriocheir sinensis.
Two Rab GTPases, EsRab-1 and EsRab-3, involved in anti-bacterial response of Chinese mitten crab Eriocheir sinensis.
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DOI：
10.1016/j.fsi.2013.06.010
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发表时间：
2013-09
影响因子：
4.7
通讯作者：
Leilei Wang;Ling Li;Lingling Wang;Jialong Yang;Jingjing Wang;Zhi Zhou;Huan Zhang;L. Song
中科院分区：
文献类型：
--
作者：
Leilei Wang;Ling Li;Lingling Wang;Jialong Yang;Jingjing Wang;Zhi Zhou;Huan Zhang;L. Song
Rab GT3在所有真核细胞中控制细胞内膜运输是必需的，并进一步影响吞噬细胞吞噬病原体的能力。本研究采用cDNA末端快速扩增（RACE）方法和表达序列标签（EST）分析，从中华绒螯蟹（Eriocheir sinensis）中鉴定了两种蟹Rab蛋白（EsRab-1和EsRab-3）的cDNA。EsRab-1和EsRab-3的cDNA全长分别为892 bp和965 bp，开放阅读框分别为615 bp和630 bp。cDNA编码204和209个氨基酸残基的两个肽段，具有保守的GTP/Mg ~（2+）结合位点，RAB结构域中的Switch I区和Switch II区。EsRab-1和EsRab-3在肝胰腺中表达量最高，在血细胞、肌肉、性腺、鳃和心脏中表达量较低。鳗弧菌攻击后，血细胞中EsRab-1和EsRab-3的表达量均显著上调，在1.5 h达到最高水平，分别是空白组的7倍（P< 0.05）和6倍（P< 0.01）。经毕赤酵母刺激后，蟹体内EsRab-1和EsRab-3的mRNA表达量均无明显变化。这些结果清楚地表明Rab蛋白参与螃蟹的抗细菌免疫。
Rab GTPase is essential for the control of intracellular membrane trafficking in all eukaryotic cells and further affects the ability of phagocytic cells to scavenge pathogen. In the present study, the cDNAs for two crab Rab proteins (EsRab-1 and EsRab-3) were identified from the Chinese mitten crabEriocheir sinensisby rapid amplification of cDNA ends (RACE) approaches and expressed sequence tag (EST) analysis. The full-length cDNAs of EsRab-1 and EsRab-3 were of 892 bp and 965 bp with ORFs of 615 bp and 630 bp, respectively. The cDNAs encoded two peptides of 204 and 209 amino acid residues with the conserved GTP/Mg2+binding sites, Switch I region and Switch II region in RAB domains. The mRNA transcripts of EsRab-1 and EsRab-3 were both highest expressed in hepatopancreas, and marginally expressed in other tissues including hemocytes, muscle, gonad, gill and heart. After the crabs were challenged by bacteria Vibrio anguillarum, the expression levels of both EsRab-1 and EsRab-3 in hemocytes were significantly up-regulated, and reached the highest level at 1.5 h post-stimulation, which was 7-fold (P< 0.05) and 6-fold (P< 0.01) of blank group for EsRab-1 and EsRab-3, respectively. No significant change of mRNA expression was detected for either EsRab-1 or EsRab-3 in crabs stimulated by Pichia pastoris. These results clearly suggested the involvement of Rab proteins in crab anti-bacterial immunity.