Laboratory of Insect Toxicology, Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, South China Agricultural University, Guangzhou, People's Republic of China.
PLoS One. 2013;8(3):e59527. doi: 10.1371/journal.pone.0059527. Epub 2013 Mar 26.
Deregulated reactive oxygen species (ROS) production can lead to the disruption of structural and functional integrity of cells as a consequence of reactive interaction between ROS and various biological components. Catalase (CAT) is a common enzyme existing in nearly all organisms exposed to oxygen, which decomposes harmful hydrogen peroxide, into water and oxygen. In this study, the full length sequence that encodes CAT-like protein from Spodoptera litura named siltCAT (GenBank accession number: JQ_663444) was cloned and characterized. Amino acid sequence alignment showed siltCAT shared relatively high conservation with other insect, especially the conserved residues which defined heme and NADPH orientation. Expression pattern analysis showed that siltCAT mRNA was mainly expressed in the fat body, midgut, cuticle and malpighian tube, and as well as over last instar larvae, pupa and adult stages. RNA interference was used to silence CAT gene in SL-1 cells and the fourth-instar stage of S. litura larvae respectively. Our results provided evidence that CAT knockdown induced ROS generation, cell cycle arrest and apoptosis in SL-1 cells. It also confirmed the decrease in survival rate because of increased ROS production in experimental groups injected with double-stranded RNA of CAT (dsCAT). This study implied that ROS scavenging by CAT is important for S. litura survival.
ROS 产生失调会导致细胞的结构和功能完整性被破坏,这是由于 ROS 与各种生物成分之间的反应性相互作用所致。CAT(过氧化氢酶)是一种存在于几乎所有接触氧气的生物体中的常见酶,它能将有害的过氧化氢分解为水和氧气。在这项研究中,我们从斜纹夜蛾中克隆并鉴定了编码 CAT 样蛋白的全长序列,命名为 siltCAT(GenBank 登录号:JQ_663444)。氨基酸序列比对显示,siltCAT 与其他昆虫具有相对较高的同源性,尤其是定义血红素和 NADPH 取向的保守残基。表达模式分析表明,siltCAT mRNA 主要在脂肪体、中肠、表皮和马氏管中表达,并且在末龄幼虫、蛹和成虫阶段也有表达。我们使用 RNA 干扰技术分别在 SL-1 细胞和斜纹夜蛾的第四龄幼虫中沉默 CAT 基因。我们的结果表明,CAT 敲低会诱导 SL-1 细胞中 ROS 的产生、细胞周期停滞和凋亡,同时还证实了由于 CAT 的双链 RNA(dsCAT)注射实验组中 ROS 产生增加,导致生存率下降。这项研究表明,CAT 通过清除 ROS 对斜纹夜蛾的生存是很重要的。