PROTEIN NUCLEIC ACID INTERACTIONS

蛋白质核酸相互作用

• 批准号: 6385585
• 负责人: PAUL J HAGERMAN
• 金额: $ 25.63万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-07-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6385585
• 关键词: DNA; DNA binding protein; RNA; RNA binding protein; bioengineering /biomedical engineering; biomedical equipment development; biophysics; birefringences; chemical stability; computer simulation; conformation; fluorescence resonance energy transfer; intermolecular interaction; method development; nucleic acid sequence; nucleic acid structure; protein binding; rare earth element; solutions; structural biology; technology /technique development; virus RNA; virus protein

The proposed research has three principal objectives: (I) a more quantitative understanding of the sequence-dependence and origins of nucleic acid helix rigidity; (ii)characterization of the conformational changes induced in DNA (or RNA) by three site- specific nucleic acid binding proteins; (iii) further development of two solution-based methods, namely, transient electric birefringence (TEB) and luminescence resonance energy transfer (LRET), for studying the long-range conformations of nucleic acids. Used in combination, these two methods should complement one another to reduce the experimental and theoretical uncertainties associated with either method alone. Each of the three protein-nucleic acid interactions is designed to examine a somewhat different issue: The TATA-box protein (TBP) is a critical component of the eukaryotic transcriptional machinery; its complex with DNA provides an excellent test system for examining large bends in solution, and for laying the groundwork for a more comprehensive study of the specific interactions that give rise to DNA bending. The basic region- leucine zipper protein (bZIP) family is extremely important in the process of tissue-specific transcriptional regulation; examination of the bZIP-DNA interaction will hopefully resolve the question of how much (or whether) those proteins bend their DNA targets. The third study will focus on the nature of the conformational change induced in a plant viral RNA genome (alfalfa mosaic virus) by site-specific binding of the viral coat protein. The RNA target is likely to possess a significant degree of flexibility prior to protein binding, thus providing a test of the TEB/LRET approach to assess flexibility.

拟议的研究具有三个主要目标：（i）对核酸螺旋刚度的序列依赖性和起源的更定量理解； （ii）三种位点特异性核酸结合蛋白在DNA（或RNA）中诱导​​的构象变化的表征； （iii）进一步开发了两种基于溶液的方法，即瞬时电力双折射（TEB）和发光谐振能量转移（LRET），用于研究核酸的远距离构象。 结合使用，这两种方法应相互补充，以减少与任何一种方法相关的实验和理论不确定性。三种蛋白质核酸相互作用中的每一个都旨在检查一个不同的问题：TATA-box蛋白（TBP）是真核转录机械的关键组成部分；它与DNA的复合物为检查溶液中的大弯曲提供了出色的测试系统，并为对引起DNA弯曲的特定相互作用的更全面研究奠定了基础。 基本区域 - 亮氨酸拉链蛋白（BZIP）家族在组织特异性的转录调控过程中非常重要。对BZIP-DNA相互作用的检查将有望解决这些蛋白质弯曲其DNA靶标的（或是否）的问题。 第三项研究将侧重于通过病毒外套蛋白的位点特异性结合植物病毒RNA基因组（苜蓿镶嵌病毒）引起的构象变化的性质。 RNA靶标在蛋白质结合之前可能具有很大程度的柔韧性，因此提供了TEB/LRET方法的测试以评估柔韧性。