Hypoxia Affects the Antioxidant Activity of Glutaredoxin 3 in <i>Scylla paramamosain</i> through Hypoxia Response Elements
Hypoxia is a major environmental stressor that can damage the oxidation metabolism of crustaceans. Glutaredoxin (Grx) is a key member of the thioredoxin superfamily and plays an important role in the host's defense against oxidative stress. At present, the role of Grx in response to hypoxia in...
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| Main Authors: | , , , , , , |
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| Format: | Book |
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MDPI AG,
2022-12-01T00:00:00Z.
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| Online Access: | Connect to this object online. |
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| Summary: | Hypoxia is a major environmental stressor that can damage the oxidation metabolism of crustaceans. Glutaredoxin (Grx) is a key member of the thioredoxin superfamily and plays an important role in the host's defense against oxidative stress. At present, the role of Grx in response to hypoxia in crustaceans remains unclear. In this study, the full-length cDNA of Grx3 (<i>SpGrx3</i>) was obtained from the mud crab <i>Scylla paramamosain</i>, which contains a 129-bp 5' untranslated region, a 981-bp open reading frame, and a 1,183-bp 3' untranslated region. The putative SpGrx3 protein contains an N-terminal thioredoxin domain and two C-terminal Grx domains. <i>SpGrx3</i> was expressed in all tissues examined, with the highest expression in the anterior gills. After hypoxia, <i>SpGrx3</i> expression was significantly up-regulated in the anterior gills of mud crabs. The expression of Grx2 and glutathione S-transferases was decreased, while the expression of glutathione peroxidases was increased following hypoxia when <i>SpGrx3</i> was silenced <i>in vivo</i>. In addition, the total antioxidant capacity of <i>SpGrx3</i>-interfered mud crabs was significantly decreased, and the malondialdehyde content was significantly increased during hypoxia. The subcellular localization data indicated that SpGrx3 was predominantly localized in the nucleus when expressed in <i>Drosophila</i> Schneider 2 (S2) cells. Moreover, overexpression of SpGrx3 reduced the content of reactive oxygen species in S2 cells during hypoxia. To further investigate the transactivation mechanism of <i>SpGrx3</i> during hypoxia, the promoter region of the <i>SpGrx3</i> was obtained by Genome Walking and three hypoxia response elements (HREs) were predicted. Dual-luciferase reporter assay results demonstrated that <i>SpGrx3</i> was likely involved in the hypoxia-inducible factor-1 (HIF-1) pathway during hypoxia, which could be mediated through HREs. The results indicated that <i>SpGrx3</i> is involved in regulating the antioxidant system of mud crabs and plays a critical role in the response to hypoxia. |
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| Item Description: | 10.3390/antiox12010076 2076-3921 |