A Litopenaeus vannamei p70S6K gene is involved in the antioxidative and apoptosis under low temperature

Abstract

p70S6K is involved in cellular response, such as tumor metastases, the immune response and tissue repair in vertebrates. The role of p70S6K in these physiological processes in crustaceans remains, however, unknown. In this study, the Lvp70S6K was identified, containing a 5' UTR of 294 bp, an ORF of 1494 bp ad a 3' UTR of 211 bp, encoding 497 amino acids with a theoretical molecular weight of 70 kDa and an estimated isoelectric point of (pI) of 5.16. The multiple alignment found that Lvp70S6K was highly homologous with other invertebrates. Lvp70S6K mRNA was detected in all the tested tissues and the Lvp70S6K expression levels was significantly down-regulated and reached the lowest level (0.44-fold, p < 0.01) at 1.5 h after low temperature stress. The subcellular localization of Lvp70S6K could be detected in cytoplasm. ROS production was significantly up-regulation (1.19-fold, p < 0.01), total hemocyte count (THC) was significantly down-regulation (0.22-fold, p < 0.01), apoptosis rate was markedly increased (1.09-fold, p < 0.01), apoptosis-related genes of LvPDCD4 (1.61-fold, p < 0.01) and LvCyt.C (1.23-fold, p < 0.01) were up-regulated, and anti-apoptotic gene of LvBcl-2 (0.69-fold, p < 0.01), LvIAP1 (0.68-fold, p < 0.01) and LvIAP2 (0.45-fold, p < 0.01) were decreased after low temperature stress in hemolymph of Lvp70S6K-silenced shrimp at 1.5 h. Silencing of LvPTEN significantly increased Lvp70S6K, LvPI3K, LvAKT and LvmTOR expression. In summary, these results indicated that Lvp70S6K play a crucial role in oxidative and apoptosis, which was able to negatively regulate by PTEN.

Keywords: Antioxidative; Apoptosis; Litopenaeus vannamei; Low temperature stress; p70S6K.