Chan M T, Yu S M
Institute of Molecular Biology, Academia Sinica, Nankang, Taipei 11529, Taiwan, Republic of China.
Proc Natl Acad Sci U S A. 1998 May 26;95(11):6543-7. doi: 10.1073/pnas.95.11.6543.
In plants, sugar feedback regulation provides a mechanism for control of carbohydrate allocation and utilization among tissues and organs. The sugar repression of alpha-amylase gene expression in rice provides an ideal model for studying the mechanism of sugar feedback regulation. We have shown previously that sugar repression of alpha-amylase gene expression in rice suspension cells involves control of both transcription rate and mRNA stability. The alpha-amylase mRNA is significantly more stable in sucrose-starved cells than in sucrose-provided cells. To elucidate the mechanism of sugar-dependent mRNA turnover, we have examined the effect of alphaAmy3 3' untranslated region (UTR) on mRNA stability by functional analyses in transformed rice suspension cells. We found that the entire alphaAmy3 3' UTR and two of its subdomains can independently mediate sugar-dependent repression of reporter mRNA accumulation. Analysis of reporter mRNA half-lives demonstrated that the entire alphaAmy3 3' UTR and the two subdomains each functioned as a sugar-dependent destabilizing determinant in the turnover of mRNA. Nuclear run-on transcription analysis further confirmed that the alphaAmy3 3' UTR and the two subdomains did not affect the transcription rate of promoter. The identification of sequence elements in the alpha-amylase mRNA that dictate the differential stability has very important implications for the study of sugar-dependent mRNA decay mechanisms.
在植物中,糖反馈调节为控制碳水化合物在组织和器官间的分配与利用提供了一种机制。水稻中α-淀粉酶基因表达的糖抑制作用为研究糖反馈调节机制提供了一个理想模型。我们之前已经表明,水稻悬浮细胞中α-淀粉酶基因表达的糖抑制涉及转录速率和mRNA稳定性的控制。α-淀粉酶mRNA在蔗糖饥饿的细胞中比在提供蔗糖的细胞中显著更稳定。为了阐明糖依赖性mRNA周转的机制,我们通过在转化的水稻悬浮细胞中的功能分析,研究了αAmy3 3'非翻译区(UTR)对mRNA稳定性的影响。我们发现整个αAmy3 3' UTR及其两个亚结构域可以独立介导报告基因mRNA积累的糖依赖性抑制。对报告基因mRNA半衰期的分析表明,整个αAmy3 3' UTR和两个亚结构域在mRNA周转中均作为糖依赖性去稳定决定因素发挥作用。细胞核连续转录分析进一步证实,αAmy3 3' UTR和两个亚结构域不影响启动子的转录速率。鉴定α-淀粉酶mRNA中决定差异稳定性的序列元件对研究糖依赖性mRNA降解机制具有非常重要的意义。