Transcript profiles of mitochondrial and cytoplasmic manganese superoxide dismutases in Exopalaemon carinicauda under ammonia stress

• Published in: Chinese journal of oceanology and limnology Vol. 33; no. 3; pp. 714 - 724
• Main Author: 任海 李健 李吉涛 刘萍 梁忠秀 吴建华
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer-Verlag 01.05.2015; Science Press; Springer Nature B.V
• Subjects: 3' Untranslated regions; 5' Untranslated Regions; Amino acid sequence; Amino acid sequences; Amino acids; Ammonia; Antioxidants; Binding sites; Biology; Callinectes sapidus; cDNA末端; Cloning; Complementary DNA; Danio rerio; Earth and Environmental Science; Earth Sciences; Environmental stress; Enzymes; Exopalaemon carinicauda; Freshwater crustaceans; Glycosylation; Hemocytes; Hepatopancreas; humans; isoelectric point; Macrobrachium nipponense; Macrobrachium rosenbergii; Manganese; Marine crustaceans; Molecular weight; Nucleotide sequence; Oceanography; open reading frames; Oxidative stress; PCR; Perisesarma bidens; Proteins; rapid amplification of cDNA ends; reverse transcriptase polymerase chain reaction; RT-PCR分析; sequence analysis; Sequencing; Shellfish; shrimp; signal peptide; Stress response; Superoxide dismutase; tissues; 成绩; 氨基酸序列; 线粒体; 细胞质; 脊尾白虾; 锰超氧化物歧化酶; ); expression; cytosolic manganese superoxide dismutase; mitochondrial manganese superoxide dismutase; cloning
• ISSN: 0254-4059; 2096-5508; 1993-5005; 2523-3521
• DOI: 10.1007/s00343-015-4143-5

Superoxide dismutase （SOD） is one of the most important antioxidant defense enzymes, and is considered as the first line against oxidative stress. In this study, we cloned a mitochondrial manganese （Mn） SOD （mMnSOD） cDNA from the ridgetail white prawn Exopalaemon carinicauda by using rapid amplification of cDNA ends （RACE） methods. The fulMength cDNA for mMnSOD was 1 014-bp long, containing a 5＇-untranslated region （UTR） of 37-bp, a 3＇-UTR of 321-bp with a poly （A） tail, and included a 657-bp open reading frame encoding a protein of 218 amino acids with a 16-amino-acid signal peptide. The protein had a calculated molecular weight of 23.87 kDa and a theoretical isoelectric point of 6.75. The mMnSOD sequence included two putative N-glycosylation sites （NHT and NLS）, the MnSOD signature sequence 18~DVWEHAYY~87, and four putative Mn binding sites （H48, H96, D180, and H184）. Sequence comparison showed that the mMnSOD deduced amino acid sequence of E. carinicauda shared 97%, 95%, 89%, 84%, 82%, 72%, and 69% identity with that ofMacrobrachium rosenbergii, Macrobrachium nipponense, Fenneropeneaus chinensis, Callinectes sapidus, Perisesarma bidens, Danio rerio, and Homo sapiens, resectively. Quantitative real-time RT-PCR analysis showed that mMnSOD transcripts were present in all E. carinicauda tissues examined, with the highest levels in the hepatopancreas. During an ammonia stress treatment, the transcript levels of mMnSOD and cMnSOD were up-regulated at 12 h in hemocytes and at 24 h in the hepatopancreas. As the duration of the ammonia stress treatment extended to 72 h, the transcript levels of mMnSOD and cMnSOD significantly decreased both in hemocytes and hepatopancreas. These findings indicate that the SOD system is induced to respond to acute ammonia stress, and may be involved in environmental stress responses in E. carinicauda.