Abstract To assure an efficient protein production under stress conditions, some transcripts are modulated at the translational level. To better understand the control of protein production under stress, we analysed the enhancing effect of the 3’ untranslated region (UTR) of the water deficit responsive gene PvLEA6 from Phaseolus vulgaris on the regulation of its expression. GUS activity and transcript loading on polysomes were analysed using Arabidopsis transgenic plants with constructs containing GUS open reading frame fused to wild type or mutated PvLEA6-3’UTR, driven by the PvLEA6 promoter, grown under optimal or stress conditions. The effect of the PvLEA6-3’UTR on gene transcription and transcript stability was discarded. We demonstrated that the PvLEA6-3’UTR allows preferential polysome loading of the GUS reporter transcript under water deficit. The effect of this region on protein synthesis was also supported by in vitro translation data. Particular conserved motifs in this region responsible for translation stimulation were identified. Specific interaction of the 3’UTR with cellular protein(s) was shown. Our data support a specific role of PvLEA6-3’UTR leading to efficient protein synthesis and hence a competent response under water deficit and suggest the participation of mRNA binding proteins in translational enhancement of the PvLEA6 mRNA.

中文翻译：

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由编码第 6 组晚期胚胎发生丰富蛋白的转录本的 3' 非翻译区赋予的翻译增强

摘要 为了确保在压力条件下有效的蛋白质生产，一些转录本在翻译水平上进行了调节。为了更好地了解胁迫下蛋白质生产的控制，我们分析了来自菜豆的缺水响应基因 PvLEA6 的 3' 非翻译区 (UTR) 对其表达调控的增强作用。使用拟南芥转基因植物分析 GUS 活性和多核糖体上的转录物加载，其构建体包含 GUS 开放阅读框，融合到野生型或突变的 PvLEA6-3'UTR，由 PvLEA6 启动子驱动，在最佳或胁迫条件下生长。PvLEA6-3'UTR 对基因转录和转录稳定性的影响被丢弃。我们证明了 PvLEA6-3'UTR 允许在缺水情况下优先加载 GUS 报告基因转录本。该区域对蛋白质合成的影响也得到了体外翻译数据的支持。鉴定了该区域中负责翻译刺激的特定保守基序。显示了 3'UTR 与细胞蛋白质的特异性相互作用。我们的数据支持 PvLEA6-3'UTR 的特定作用，导致有效的蛋白质合成，从而在缺水情况下产生有效的反应，并表明 mRNA 结合蛋白参与 PvLEA6 mRNA 的翻译增强。显示了 3'UTR 与细胞蛋白质的特异性相互作用。我们的数据支持 PvLEA6-3'UTR 的特定作用，导致有效的蛋白质合成，因此在缺水情况下产生有效的反应，并表明 mRNA 结合蛋白参与 PvLEA6 mRNA 的翻译增强。显示了 3'UTR 与细胞蛋白质的特异性相互作用。我们的数据支持 PvLEA6-3'UTR 的特定作用，导致有效的蛋白质合成，从而在缺水情况下产生有效的反应，并表明 mRNA 结合蛋白参与 PvLEA6 mRNA 的翻译增强。