Bioorganic Basis for RNA-Protein Stability

RNA-蛋白质稳定性的生物有机基础

• 批准号: 6768798
• 负责人: ANNE M BARANGER
• 金额: $ 28.3万
• 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/6768798
• 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; phage display; 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-蛋白质复合体是许多重要生物过程的重要贡献者，包括基因表达。了解蛋白质如何识别具有高亲和力和高特异性的RNA，对于描述和控制这些过程将是有价值的。这项研究的目标是确定RNA-蛋白质复合体形成的能量贡献者的特征。拟议的研究将集中在RNA识别基序(RRM)对RNA的识别上，RRM是最常见的RNA结合域之一。在上一个资助期间，参与与RNA碱基堆积相互作用的高度保守的芳香氨基酸被发现对U1A-RNA复合体的稳定性有特殊贡献。在目前的资助期间，这些保守的芳香族氨基酸提供稳定的分子来源将被调查。通过将蛋白质突变和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相结合，探讨它们对复合体中的堆积相互作用、对游离蛋白质和复合体的结构以及结合所需的构象变化的贡献。为了探索用U1A获得的结果的普遍性，将对其他RRM-RNA复合物进行类似的研究。比较不同RRM-RNA复合体中的结合将有助于识别RRM-RNA复合体中共同持有的能量贡献。最后，RRM将被用作模板，利用噬菌体展示技术开发与RNA结合的小β发夹多肽。特别是，芳香氨基酸稳定β-发夹-RNA相互作用的能力将被研究。总之，这项拟议的研究将揭示芳香氨基酸如何控制RNA-蛋白质相互作用的新模型。这些模型在开发选择性控制涉及RNA-蛋白质复合体的生物过程的工具方面应该是有用的。