BIOORGANIC BASIS FOR RNA/PROTEIN STABILITY

RNA/蛋白质稳定性的生物有机基础

• 批准号: 6342975
• 负责人: ANNE M BARANGER
• 金额: $ 16.56万
• 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 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6342975
• 关键词: RNA; RNA binding protein; chemical bond; chemical stability; chemical structure function; hydrogen bond; intermolecular interaction; molecular shape; peptide library; structural biology

DESCRIPTION: The long-term goal of this research program is to understand the molecular mechanism of RNA-protein recognition. The proposed research will investigate the role of stacking interactions in stabilizing RNA-protein complexes. Stacking interactions have been proposed to be a general mechanism by which proteins specifically recognize RNA, yet little is known about their energetic contribution to RNA-protein complex formation. A quantitative description of the contribution of stacking interactions to RNA-protein binding is essential for applications that seek to modify or control processes involving RNA. Experiments will focus on the RNP domain, one of the most prevalent RNA-binding domains. Proteins containing RNP domains are important in developmental pathways and are implicated in autoimmune diseases. RNP domains have three, well conserved aromatic amino acids in the RNA-binding region that are observed to participate in stacking interactions with RNA bases in structural studies. In order to probe the electronic factors that contribute to stacking interactions in RNA-RNA complexes, the aromatic residues will be systematically varied through substitutions of natural and unnatural amino acids. Small molecule model studies will be used to evaluate the stacking ability of the amino acid side chains. A correlation between the stacking ability of the amino acid side chain and the equilibrium dissociation constants of the RNA-RNA complexes will demonstrate that stacking interactions are important for binding. To probe cooperation between hydrogen bonding and stacking interactions in RNA-RNA complexes, individual hydrogen bonds will be eliminated in modified bases that are incorporated in the RNA binding site. The ability of proteins to use stacking interactions as a means to distinguish between RNA and DNA binding sites will be probed through an investigation of stacking interactions in a DNA-peptide complex. Finally, combinatorial experiments will reveal whether stacking interactions are required for RNA recognition by the RNP domain.

描述：本研究计划的长期目标是了解 RNA-蛋白质识别的分子机制。 拟议研究 将研究堆叠相互作用在稳定 RNA-蛋白质复合物。 堆叠相互作用被认为是一种 蛋白质特异性识别RNA的一般机制，但很少 已知它们对RNA-蛋白质复合物能量贡献 阵 叠加作用的定量描述 RNA-蛋白质结合的相互作用对于寻求 修饰或控制RNA的过程。 实验将集中在 RNP结构域是最常见的RNA结合结构域之一。 蛋白 含有RNP结构域的蛋白质在发育途径中很重要， 与自身免疫性疾病有关 RNP结构域具有三个高度保守的 RNA结合区中的芳香族氨基酸被观察到， 在结构研究中参与与RNA碱基的堆积相互作用。 为了探测有助于堆积的电子因素 在RNA-RNA复合物中的相互作用，芳香残基将被 通过天然和非天然氨基的取代系统地改变 acids. 小分子模型研究将用于评估堆叠 氨基酸侧链的能力。 一个相互关联的堆积 氨基酸侧链的能力和平衡解离 RNA-RNA复合物的常数将证明， 相互作用对于结合是重要的。 探讨合作， RNA-RNA复合物中的氢键和堆积相互作用，个体 掺入的修饰碱基中的氢键将被消除 RNA结合位点。 蛋白质利用堆积相互作用的能力 作为区分RNA和DNA结合位点的手段， 通过研究DNA-肽复合物中的堆积相互作用。 最后，组合实验将揭示堆叠相互作用是否 是RNP结构域识别RNA所必需的。