Characterization and expression of cytoplasmic copper/zinc superoxide dismutase (CuZn SOD) gene under temperature and hydrogen peroxide (H2O2) in rotifer Brachionus calyciflorus

• Published in: Gene Vol. 518; no. 2; pp. 388 - 396
• Main Authors: Yang, Jianghua, Dong, Siming, Jiang, Qichen, Si, Qin, Liu, Xuezhou, Yang, Jiaxin
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.04.2013
• Subjects: Amino Acid Sequence; amino acid sequences; amino acids; Animals; antioxidants; Base Sequence; Brachionus calyciflorus; Caenorhabditis elegans; Caenorhabditis elegans - enzymology; Caenorhabditis elegans - genetics; Carps - genetics; catalase; Catalase - genetics; Cloning, Molecular; complementary DNA; Copper/zinc superoxide dismutase; DNA, Complementary; Drosophila melanogaster; Drosophila melanogaster - enzymology; Drosophila melanogaster - genetics; gene expression; gene expression regulation; genes; Hydrogen Peroxide; messenger RNA; Molecular Sequence Data; open reading frames; Phylogeny; RNA, Messenger - genetics; RNA, Messenger - metabolism; Rotifera - enzymology; Rotifera - genetics; Schistosoma japonicum; Schistosoma japonicum - enzymology; Schistosoma japonicum - genetics; Sequence Analysis, DNA; superoxide dismutase; Superoxide Dismutase - chemistry; Superoxide Dismutase - genetics; Superoxide Dismutase - metabolism; Temperature; Vitamin E; Copper/zinc superoxide dismutase; ORF; RNAi; Temperature; O2; Hydrogen peroxide; GST; SOD; β-actin; Mn SOD; GR; BBM; H2O2; GPx; CR; Ct; Vitamin E; RT-PCR; CAT; CuZn SOD; EPA; ROS; Brachionus calyciflorus; GSH
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2012.12.101

Superoxide dismutase (SOD, EC 1.15.1.1) is an important antioxidant enzyme that protects organs from damage by reactive oxygen species (ROS). We cloned cDNA encoding SOD activated with copper/zinc (CuZn SOD) from the rotifer Brachionus calyciflorus Pallas. The full-length cDNA of CuZn SOD was 692bp and had a 465bp open reading frame encoding 154 amino acids. The deduced amino acid sequence of B. calyciflorus CuZn SOD showed 63.87%, 60.00%, 59.74% and 48.89% similarity with the CuZn SOD of the Ctenopharyn godonidella, Schistosoma japonicum, Drosophila melanogaster and Caenorhabditis elegans, respectively. The phylogenetic tree constructed based on the amino acid sequences of CuZn SODs from B. calyciflorus and other organisms revealed that rotifer is closely related to nematode. Analysis of the expression of CuZn SOD under different temperatures (15, 30 and 37°C) revealed that its expression was enhanced 4.2-fold (p<0.001) at 30°C after 2h, however, the lower temperature (15°C) promoted CuZn SOD transiently (4.1-fold, p<0.001) and then the expression of CuZn SOD decreased to normal level (p>0.05). When exposed to H2O2 (0.1mM), CuZn SOD, manganese superoxide dismutase (Mn SOD) and catalase (CAT) gene were upregulated, and in addition, the mRNA expression of CuZn SOD gene was induced instantaneously after exposure to vitamin E. It indicates that the CuZn SOD gene would be an important gene in response to oxidative and temperature stress. ► We cloned cytoplasmic CuZn SOD gene from rotifer Brachionus calyciflorus. ► Characterized the B. calyciflorus CuZn SOD gene. ► Analyzed the changes of CuZn SOD expression under temperature stress and H2O2. ► Studied the effect of vitamin E on CuZn SOD, Mn SOD and CAT gene expression.