Bioorganic Basis for RNA-Protein Stability

RNA-蛋白质稳定性的生物有机基础

• 批准号: 6683264
• 负责人: ANNE M BARANGER
• 金额: $ 31.8万
• 依托单位: WESLEYAN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6683264
• 关键词: RNA; RNA binding protein; X ray crystallography; aminoacid; binding sites; bioenergetics; chemical bond; chemical kinetics; chemical stability; chemical structure function; computer simulation; fluorescence spectrometry; gene expression; genetic manipulation; hydrogen bond; intermolecular interaction; molecular dynamics; molecular shape; nuclear magnetic resonance spectroscopy; peptide library; protein binding; structural biology; thermodynamics

DESCRIPTION (provided by applicant): RNA-protein complexes are essential contributors to many important biological processes, including gene expression. An understanding of how proteins recognize RNA with high affinity and specificity will be valuable for describing and controlling these processes. The goal of this research program is to characterize the energetic contributors to RNA-protein complex formation. The proposed research will focus on the recognition of RNA by the RNA recognition motif (RRM), which is one of the most common RNA-binding domains. In the previous funding period, highly conserved aromatic amino acids that are involved in stacking interactions with RNA bases were found to be exceptional contributors to the stability of the U1A-RNA complex. In the current funding period, the molecular origins of the stabilization provided by these conserved aromatic amino acids will be investigated. Their contributions to stacking interactions in the complex, to the structures of the free protein and the complex, and to the conformational changes required upon binding will be probed by combining protein mutagenesis and synthetic modification of the RNA with structural studies, binding thermodynamic and kinetic measurements, and molecular dynamics simulations. To probe the generality of the results obtained with U1A, similar investigations with other RRM-RNA complexes will be performed. The comparison of binding in different RRM-RNA complexes will enable the identification of the energetic contributions that are held in common in RRM-RNA complexes. Finally, the RRM will be used as a template to develop small beta-hairpin peptides that bind RNA using phage display techniques. In particular, the ability of aromatic amino acids to stabilize beta-hairpin-RNA interactions will be investigated. Together, the proposed research will uncover new models for how aromatic amino acids control RNA-protein interactions. These models should be useful in the development of tools for the selective control of biological processes involving RNA-protein complexes.

描述（由申请人提供）：RNA蛋白质复合物是许多重要的生物学过程的重要因素，包括基因表达。了解蛋白质如何识别具有高亲和力和特异性的RNA对于描述和控制这些过程将是有价值的。该研究计划的目的是表征RNA-蛋白质复合物形成的能量贡献者。拟议的研究将集中于RNA识别基序（RRM）对RNA的识别，RNA识别基序是最常见的RNA结合域之一。在上一个资金期间，发现与RNA碱基堆叠相互作用的高度保守的芳族氨基酸是U1A-RNA复合物稳定性的非凡贡献者。在当前的资金期间，将研究这些保守的芳香族氨基酸提供的稳定性的分子起源。它们对复合物中堆叠相互作用的贡献，对自由蛋白和复合物的结构以及结合后所需的构象变化的贡献，将通过将蛋白质诱变和RNA的合成修饰与结构研究，结合热力学和动力学测量以及分子动力学模拟结合，结合RNA进行探测。为了探测用U1A获得的结果的一般性，将对其他RRM-RNA复合物进行类似的研究。在不同的RRM-RNA复合物中结合的比较将使能够鉴定在RRM-RNA复合物中具有共同的能量贡献。最后，RRM将用作模板，以开发使用噬菌体显示技术结合RNA的小β-发射肽。特别是，将研究芳香族氨基酸稳定β-发射β-RNA相互作用的能力。拟议的研究将共同​​发现芳族氨基酸如何控制RNA-蛋白质相互作用的新模型。这些模型应该在开发涉及RNA-蛋白质复合物的生物过程的工具的开发中有用。