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

Abstract

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 °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 ZnSO₄ 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.

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