Chan M T, Yu S M
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, Republic of China.
Plant J. 1998 Sep;15(5):685-95. doi: 10.1046/j.1365-313x.1998.00244.x.
A decrease in transcript stability is one of the important mechanisms that control the sugar repression of alpha-amylase gene expression in rice suspension cells. In this study, we investigated the function of the 3' untranslated region (3'UTR) of a rice alpha-amylase gene, alpha Amy3, in relation to sugar-dependent accumulation of mRNA. By examining the transient expression of chimeric genes in rice protoplasts, we were able to demonstrate that the alpha Amy3 3'UTR mediated the sugar-dependent repression of fused heterologous gene expression. The same kinetics of accumulation of alpha Amy3 mRNA and reporter mRNA carrying the alpha Amy3 3'UTR in response to glucose deprivation were observed, suggesting that the alpha Amy3 3'UTR is sufficient, and probably the major determinant for controlling the abundance of these transcripts. Functional analysis of two subdomains of alpha Amy3 3'UTR by insertion into a sugar-inducible chimeric gene confirmed their roles in sugar repressibility. The regulatory sequences in the alpha Amy3 3'UTR may act as potent determinants of mRNA stability in response to sugar availability. This finding has important implications for studying the regulatory mechanism of sugar repression in eukaryotes.
转录本稳定性的降低是控制水稻悬浮细胞中α-淀粉酶基因表达的糖抑制作用的重要机制之一。在本研究中,我们研究了水稻α-淀粉酶基因α Amy3的3'非翻译区(3'UTR)与mRNA糖依赖性积累相关的功能。通过检测水稻原生质体中嵌合基因的瞬时表达,我们能够证明α Amy3 3'UTR介导了融合异源基因表达的糖依赖性抑制。观察到α Amy3 mRNA和携带α Amy3 3'UTR的报告基因mRNA在葡萄糖剥夺时具有相同的积累动力学,这表明α Amy3 3'UTR足以,并且可能是控制这些转录本丰度的主要决定因素。通过插入糖诱导嵌合基因对α Amy3 3'UTR的两个亚结构域进行功能分析,证实了它们在糖抑制性方面的作用。α Amy3 3'UTR中的调控序列可能作为响应糖可用性的mRNA稳定性的有效决定因素。这一发现对于研究真核生物中糖抑制的调控机制具有重要意义。