Research on Function of Ribosomal Protein S6 Kinases, 1α and β, Based on Molecular Cloning and siRNA-Based Interference in Juvenile Blunt Snout Bream (Megalobrama amblycephala).

Gu, Jiaze; Mi, Haifeng; Ren, Mingchun; Huang, Dongyu; Aboseif, Ahmed Mohamed; Liang, Hualiang; Zhang, Lu

Simple Summary: The aim of this study was to investigate the effects of S6K1α and β on the expression of glycolysis- and gluconeogenesis-related genes in juvenile blunt snout bream. The two isoforms, α and β, of S6K1 in blunt snout bream were successfully cloned and characterized, and their expression patterns were examined in vivo. In this study, we designed multiple sets of siRNAs to specifically degrade s6k1α and s6k1β expression in this fish. α-siRNA inhibited both s6k1α and s6k1β expression, but β-siRNA exclusively inhibited s6k1α expression. S6K1α was more intimately involved in the regulation of gluconeogenesis when only S6K1α was inhibited, whereas the inhibition of both S6K1α and S6K1β collectively co-regulated glycolysis. The aim of this study was to investigate the effects of S6K1α and β on the expression of glycolysis- and gluconeogenesis-related genes in juvenile blunt snout bream. Two isoforms, α and β, of ribosomal protein S6 kinase 1 in blunt snout bream were cloned and characterized, and their expression patterns were examined in vivo. The sequence analysis showed that s6k1α and s6k1β contain open reading frames of 2217 and 1497 bp, encoding 738 and 498 amino acids, respectively. Both S6K1α and S6K1β consist of an S_TKc domain and an extended S_TK_X domain. s6k1α and s6k1β were abundantly expressed in the heart and gonads. siRNAs were designed, and the experiment showed that α-siRNA inhibited s6k1α and s6k1β expression, but β-siRNA exclusively inhibited s6k1α expression (p < 0.05). α-siRNA upregulated the expression levels of gk and pk, while β-siRNA upregulated pepck and g6p expression (p < 0.05). The expression of g6pdh was found to be downregulated, but the gs mRNA level was overexpressed after treatment with α-siRNA and β-siRNA (p < 0.05). In the present experiment, S6K1α was more intimately involved in the regulation of gluconeogenesis when only S6K1α was inhibited, whereas the inhibition of both S6K1α and S6K1β collectively co-regulated glycolysis.

GENE expression; MOLECULAR cloning; RNA interference; SMALL interfering RNA; PROTEIN kinases

Biology (2079-7737), 2024, Vol 13, Issue 11, p875

2079-7737

Academic Journal

10.3390/biology13110875