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)

文献类型： 外文期刊

第一作者： Gu, Jiaze

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

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关键词： molecular cloning; juvenile blunt snout bream; ribosomal protein S6 Kinase 1; RNA interference

期刊名称：BIOLOGY-BASEL （ 影响因子：3.5； 五年影响因子：4.0 ）

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年卷期： 2024 年 13 卷 11 期

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收录情况： SCI

摘要： The aim of this study was to investigate the effects of S6K1 alpha and beta on the expression of glycolysis- and gluconeogenesis-related genes in juvenile blunt snout bream. The two isoforms, alpha and beta, 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 s6k1 alpha and s6k1 beta expression in this fish. alpha-siRNA inhibited both s6k1 alpha and s6k1 beta expression, but beta-siRNA exclusively inhibited s6k1 alpha expression. S6K1 alpha was more intimately involved in the regulation of gluconeogenesis when only S6K1 alpha was inhibited, whereas the inhibition of both S6K1 alpha and S6K1 beta collectively co-regulated glycolysis. The aim of this study was to investigate the effects of S6K1 alpha and beta on the expression of glycolysis- and gluconeogenesis-related genes in juvenile blunt snout bream. Two isoforms, alpha and beta, 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 s6k1 alpha and s6k1 beta contain open reading frames of 2217 and 1497 bp, encoding 738 and 498 amino acids, respectively. Both S6K1 alpha and S6K1 beta consist of an S_TKc domain and an extended S_TK_X domain. s6k1 alpha and s6k1 beta were abundantly expressed in the heart and gonads. siRNAs were designed, and the experiment showed that alpha-siRNA inhibited s6k1 alpha and s6k1 beta expression, but beta-siRNA exclusively inhibited s6k1 alpha expression (p < 0.05). alpha-siRNA upregulated the expression levels of gk and pk, while beta-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 alpha-siRNA and beta-siRNA (p < 0.05). In the present experiment, S6K1 alpha was more intimately involved in the regulation of gluconeogenesis when only S6K1 alpha was inhibited, whereas the inhibition of both S6K1 alpha and S6K1 beta collectively co-regulated glycolysis.

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