HuR蛋白介导的人类mRNA出核调控过程的结构生物学研究

项目名称： HuR蛋白介导的人类mRNA出核调控过程的结构生物学研究

项目编号： No.31270014

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 李旭

作者单位： 中国科学技术大学

项目金额： 80万元

中文摘要： 经过两年多的研究探索，我们已完成人类前体mRNA剪切因子Im(CF Im)25KDa亚基与68KDa亚基RRM结构域蛋白质-蛋白质复合物晶体结构的解析；获得了CF Im 复合物与RNA的2.9?分辨率三元复合物结构；阐明了人类CF Im复合物识别前体mRNA“UGUAA”序列的分子机制；相关研究结果已发表在2011年的Cell Research杂志上。下一阶段我们拟针对完成3′端剪切加工修饰后的成熟mRNA的出核机制展开研究，通过解析HuR(RRM1/RRM2)-poly(U)、HuR(RRM3)-poly(A)等蛋白-RNA复合物及HuR-Trn2、HuR-PP32-Crm1、HuR(N226)-Trn2等蛋白-蛋白复合物结构，阐明该过程各效应因子间的相互作用关系，结合细胞功能实验揭示该过程分子运作机理。上述工作的开展将大大增强我们对真核细胞瞬时应答过程在转录后水平的调控机制的理解。

中文关键词： HuR蛋白；mRNA出核转运；蛋白质-RNA相互作用关系；晶体结构；复合物

英文摘要： During the past two years, we have solved the crystal structure of human CF Im, comprising CF Im25 and the RNA recognition motif domain of CF Im68 (CF Im68RRM), and the crystal structure of the CF Im-RNA complex. These structures show that two CF Im68RRM molecules bind to the CF Im25 dimer via a novel RRM-protein interaction mode forming a heterotetramer. The RNA-bound structure shows that two UGUAA RNA sequences, with anti-parallel orientation, bind to one CF Im25-CF Im68RRM heterotetramer, providing structural basis for the mechanism by which CF Im binds two UGUAA elements within one molecule of pre-mRNA simultaneously. Point mutation and kinetic analyses demonstrate that CF Im68RRM can bind the immediately flanking upstream region of the UGUAA element, and CF Im68RRM binding significantly increases the RNA-binding affinity of the complex, suggesting that CF Im68 makes an essential contribution to pre-mRNA binding. The above results have been published in the Cell Research journal in 2011. In the next phase of our research work, we plan to focus on the structural basis and functional mechanism of the exporting process from nucleus of the mature mRNA. We would like to resolve the complex structures in the HuR mediated mRNA exporting process, such as HuR(RRM1/RRM2)-poly(U) and HuR(RRM3)-poly(A) protein-RNA compl

英文关键词： HuR (Hu antigen R)；mRNA Nuclear Export；Protein-RNA Interaction；Crystal Structure；Complex