Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology Yellow Sea Fisheries Research Institute, CAFS, Qingdao, 266071, China; Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Qingdao, 266071, China.
Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology Yellow Sea Fisheries Research Institute, CAFS, Qingdao, 266071, China; Key Lab of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Qingdao, 266071, China; College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, 201306, China.
Fish Shellfish Immunol. 2019 Jan;84:998-1006. doi: 10.1016/j.fsi.2018.11.006. Epub 2018 Nov 3.
Rac1 and Rac2, belonging to the small Rho GTPase family, play an important role during the immune responses. In this study, a Rac1 homolog (CsRac1) and a Rac2 homolog (CsRac2) were cloned from the Cynoglossus semilaevis. The full-length of CsRac1 and CsRac2 cDNA was 1219 bp and 1047 bp, respectively. Both CsRac1 and CsRac2 contain a 579 bp open reading frame (ORF) which encoding a 192 amino acids putative protein. The predicted molecular weight of CsRac1 and CsRac2 was 21.41 kDa and 21.35 kDa, and their theoretical pI was 8.50 and 7.91, respectively. Sequence analysis showed that the conserved RHO domain was detected both from amino acid of CsRac1 and CsRac2. Homologous analysis showed that CsRac1 and CsRac2 share high conservation with other counterparts from different species. The CsRac1 and CsRac2 transcript showed wide tissue distribution, in which CsRac1 and CsRac2 exhibit the highest expression level in liver and gill, respectively. The expression level of CsRac1 and CsRac2 fluctuated in the liver and gill tissues at different time points after challenged by Vibrio harveyi. Specifically, CsRac1 and CsRac2 were significantly up-regulated at 48 h and 96 h post injection. Moreover, the knocking down of CsRac1 and CsRac2 in cell line (TSHKC) reduced the expression of CsPAK1, CsIL1-β and CsTNF-α. The present data suggests that CsRac1 and CsRac2 might play important roles in the innate immunity of half-smooth tongue sole.
Rac1 和 Rac2 属于小 Rho GTPase 家族，在免疫反应中发挥重要作用。本研究从半滑舌鳎中克隆得到 Rac1 同源物（CsRac1）和 Rac2 同源物（CsRac2）。CsRac1 和 CsRac2 的全长 cDNA 分别为 1219bp 和 1047bp，均含有一个 579bp 的开放阅读框（ORF），编码 192 个氨基酸的推测蛋白。CsRac1 和 CsRac2 的预测分子量分别为 21.41kDa 和 21.35kDa，理论等电点分别为 8.50 和 7.91。序列分析表明，在 CsRac1 和 CsRac2 的氨基酸中均检测到保守的 RHO 结构域。同源分析表明，CsRac1 和 CsRac2 与来自不同物种的其他同源物具有高度保守性。CsRac1 和 CsRac2 的转录本在广泛的组织中表达，其中 CsRac1 和 CsRac2 在肝脏和鳃中表达水平最高。在受到哈维弧菌刺激后不同时间点的肝脏和鳃组织中，CsRac1 和 CsRac2 的表达水平波动。具体而言，CsRac1 和 CsRac2 在注射后 48h 和 96h 时显著上调。此外，在细胞系（TSHKC）中敲低 CsRac1 和 CsRac2 降低了 CsPAK1、CsIL1-β 和 CsTNF-α 的表达。本研究数据表明，CsRac1 和 CsRac2 可能在半滑舌鳎的先天免疫中发挥重要作用。
