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 -蛋白复合物是许多重要生物过程的重要贡献者，包括基因表达。了解蛋白质如何以高亲和力和特异性识别RNA将对描述和控制这些过程有价值。本研究计划的目标是表征rna -蛋白质复合物形成的能量贡献者。RNA识别基序（RNA recognition motif， RRM）是最常见的RNA结合结构域之一，本文将重点研究RRM对RNA的识别。在之前的资助期内，高度保守的芳香族氨基酸参与了与RNA碱基的堆叠相互作用，被发现对U1A-RNA复合物的稳定性有特殊的贡献。在目前的资助期内，将研究这些保守芳香氨基酸提供稳定的分子起源。通过结合蛋白质诱变和RNA的合成修饰，结合结构研究、结合热力学和动力学测量以及分子动力学模拟，探讨它们对复合物中堆叠相互作用、自由蛋白和复合物的结构以及结合所需的构象变化的贡献。为了探究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。通过比较不同RRM-RNA复合物的结合情况，可以识别出RRM-RNA复合物中共同的能量贡献。最后，RRM将用作模板，利用噬菌体展示技术开发结合RNA的小β -发夹肽。特别是，芳香族氨基酸稳定-发夹- rna相互作用的能力将被研究。总之，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。这些模型在开发涉及rna -蛋白复合物的生物过程的选择性控制工具方面应该是有用的。