4.7 Article

Four alternative splicing transcripts of intracellular copper/zinc superoxide dismutase 1 in Oxya chinensis

Journal

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Volume 193, Issue -, Pages 1600-1609

Publisher

ELSEVIER
DOI: 10.1016/j.ijbiomac.2021.10.222

Keywords

Intracellular copper/zinc superoxide dismutase; Alternative RNA splicing; Functional analysis

Funding

  1. National Natural Science Foundation of China-China [31201548]
  2. Natural Science Foundation of Shanxi Province-China [2013011028-3]
  3. Research Fund for the Doctoral Program of Higher Education of China-China [2013M530180]

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In this study, four alternative splicing transcripts of icCuZnSOD1 in Oxya chinensis were identified, showing differences in heat resistance and antioxidant capacity. These variants exhibited varied expression patterns during developmental stages and in different tissues, with distinct responses to CuCl2 and ZnSO4. The transcripts played roles in defense against CdCl2-induced oxidative stress, with OcicCuZnSOD1a exhibiting the strongest antioxidant capacity.
In this study, we obtained four alternative splicing transcripts of intracellular copper/zinc superoxide dismutase 1 (icCuZnSOD1) in Oxya chinensis. OcicCuZnSOD1a has all common characteristics of CuZnSOD family and is a canonical CuZnSOD. OcicCuZnSOD1b is missing a Zn binding site. OcicCuZnSOD1c lacks Zn ion and is a Cu-only SOD. OcicCuZnSOD1d is missing a CuZnSOD conserved sequence and lacks the E-loop, a conserved disulfide bond, and an active site arginine. OcicCuZnSOD1a was the most heat-resistant and OcicCuZnSOD1c was the most unstable at high temperatures above 55 degrees C. They were stable at a wide pH range, especially in alkaline conditions. The four variants expressed at the throughout developmental stages and had various tissue expression patterns. OcicCuZnSOD1a and OcicCuZnSOD1d were significantly induced by 8.79 mM CuCl2 and OcicCuZnSOD1b was significantly up-regulated by 14.67 mM CuCl2. OcicCuZnSOD1a was significantly inhibited by 19.13 mM Zn50 4 while OcicCuZnSOD1d were significantly induced by 22.61 mM ZnSO4. Disc diffusion assay showed that the four isoforms of OcicCuZnSOD1 made the killing zones smaller surrounding the CdCl2-soaked filter discs. However, the reduction ratios of OcicCuZnSOD1a were the highest. These results implied that the four transcripts played roles in defense against CdCl2-induced oxidative stress while OcicCuZnSOD1a had stronger antioxidant capacity.

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