1. RNAi-mediated knockdown of catalase causes cell cycle arrest in SL-1 cells and results in low survival rate of Spodoptera litura (Fabricius)
- Author
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Hai-Ming Zhao, Liang Gong, Xiaolin Dong, Zhen Hu, Shaohua Chen, Meiying Hu, Rizwan-ul-Haq Muhammad, and Xin Yi
- Subjects
Male ,Anatomy and Physiology ,Intracellular Space ,lcsh:Medicine ,Spodoptera litura ,Gene Expression ,Apoptosis ,Biochemistry ,Antioxidants ,RNA interference ,Molecular Cell Biology ,Gene Knockdown Techniques ,Cloning, Molecular ,lcsh:Science ,chemistry.chemical_classification ,Gene knockdown ,Multidisciplinary ,Cell Death ,Gene Expression Regulation, Developmental ,Agriculture ,Catalase ,Oxygen Metabolism ,Chemistry ,Organ Specificity ,Female ,RNA Interference ,Sequence Analysis ,Research Article ,Molecular Sequence Data ,Biology ,Spodoptera ,Insect Control ,Cell Line ,Molecular Genetics ,Integrated Control ,Chemical Biology ,Genetics ,Animals ,Amino Acid Sequence ,RNA, Messenger ,RNA, Double-Stranded ,Reactive oxygen species ,lcsh:R ,fungi ,RNA ,Computational Biology ,Cell Cycle Checkpoints ,biology.organism_classification ,Molecular biology ,Survival Analysis ,Metabolism ,chemistry ,biology.protein ,lcsh:Q ,Pest Control ,Reactive Oxygen Species ,Physiological Processes ,Energy Metabolism - Abstract
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.
- Published
- 2012