野生番茄SpRing 3'UTR顺式元件及互作RNA结合蛋白的鉴定与功能验证

The 3'untranslated regions (3'UTRs) and RNA-binding proteins (RBPs) play an important role in post-transcriptional regulation of genes. So far, no 3'UTR cis-elements and RBPs involved in abiotic stress has been characterized, and this limited our understanding of their roles in signaling transduction in stress. We previously found the stability of SpRing transcripts in wild tomato species could be enhanced by its 3’UTR under salt stress conditions, which ultimately affected the expression level of SpRing. In this project, we propose to characterize the key cis-elements in 3'UTR of SpRing, and examine the involvement of 3'UTR in signaling transduction at post-transcriptional level in abiotic stress. Second, with the MS2-system and Dynabeads® Streptavidin method, we will identify the RNA-binding proteins (RBPs) that interacts with the 3’UTR of SpRing in vivo and further characterize their function. This study will provide new insights in salt stress response in plants, and prove instrumental in uncovering novel salt stress signaling pathways based on 3’UTR cis-element and RBPs.

3'UTR顺式元件与RNA结合蛋白在基因转录后调控中起关键作用。目前，在植物中尚未鉴定到特异响应逆境胁迫的3'UTR顺式元件及与其直接互作的RNA结合蛋白，极大地限制了我们对3'UTR介导胁迫信号转导的认识。在研究野生醋栗番茄耐盐基因SpRing时，我们发现SpRing基因3'UTR能特异响应盐信号，提高mRNA转录后的稳定性，最终影响SpRing的表达水平。本项目主要解决两个关键问题：一是鉴定SpRing 3'UTR的核心顺式元件，为3'UTR在转录后调控基因表达及逆境信号转导中的作用提供重要依据；二是通过我们探索建立的MS2系统及链霉亲和素磁珠法，鉴定与SpRing 3'UTR互作的RNA结合蛋白，并验证其生物学功能，揭示一条新的以3'UTR顺式元件和RNA结合蛋白为“信号传递因子”的盐胁迫信号途径。