title: Genomic organization of the cytosolic manganese superoxide dismutase gene from the Pacific white shrimp, Litopenaeus vannamei, and its response to thermal stress.
abstract: Cytosolic manganese superoxide dismutase (cMnSOD) is an important antioxidant enzyme which catalyzes the conversion of superoxides to oxygen and hydrogen peroxide in several organisms. In the Pacific white shrimp, Litopenaeus vannamei, three cMnSOD genes (LvcMnSOD1-3) have previously been characterized. Here, the genomic structure of LvcMnSOD2 and its mRNA expression in response to thermal stress was examined. Analysis of the nucleotide sequence demonstrated that LvcMnSOD2 is comprised of 2392 bp spanning from the ATG translation start site to the stop codon and contains six exons interrupted by five introns. The 5' region upstream of the LvcMnSOD2 gene contains several putative regulatory elements but lacks the accepted TATA sequence. The putative transcription factor binding elements that may be involved in LvcMnSOD2 mRNA expression level include activator protein-1 (AP-1), cAMP response element binding protein (CREB), upstream stimulatory factor (USF), CAAT-enhancer binding protein (C/EBP), nuclear factor-κB (NF-κB) and heat shock regulatory element (HSE). In addition, we compared the 5' upstream sequences of the LvcMnSOD2 gene between two shrimp strains that are resistant or susceptible to Taura syndrome virus (TSV), respectively, which revealed the absence of the USF and C/EBP elements at positions -2125 and -1986, respectively, in the TSV-susceptible shrimp line. Moreover, genomic variations between the two shrimp strains were detected in some of the putative C/EBP, USF, HSE and NF-κB transcription factor binding elements. That these genomic variations might be involved in the TSV resistance as well as in stress responses remains to be evaluated. The presence of 15 putative HSEs suggests that the expression of LvcMnSOD2 is regulated under thermal stress. Here, we found that in response to a 1 or 3 h thermal stress (35 °C), the mRNA expression levels of LvcMnSOD2 were significantly increased and then gradually decreased in the recovering phase at room temperature (25 °C) to control levels by 3 h after the heat shock. Thus, the antioxidant system may be induced to protect cells from the oxidative damage caused by thermal stress. The genomic organization of LvcMnSOD2 likely provides a clue to the mechanisms that might regulate the antioxidant defense pathway in shrimps and so potentially in marine invertebrates.
authors.affiliation: Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, 254 Phayathai Road, Bangkok 10330, Thailand; Biotechnology Program, Faculty of Science, Chulalongkorn University, 254 Phayathai Road, Bangkok 10330, Thailand.
authors.forename: Suchonma
authors.initials: S
authors.lastname: Sookruksawong
authors.forename: Siriporn
authors.initials: S
authors.lastname: Pongsomboon
authors.forename: Anchalee
authors.initials: A
authors.lastname: Tassanakajon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003600:Cytosol / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D060467:Disease Resistance / Q000235:genetics*; D040461:Gene Components; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics / Q000502:physiology*; D006358:Hot Temperature / Q000009:adverse effects*; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000201:enzymology* / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013312:Stress, Physiological / Q000235:genetics / Q000502:physiology*; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D013482:Superoxide Dismutase
keywords: Cytosolic manganese superoxide dismutase; Genomic organization; Litopenaeus vannamei; Thermal stress