STRUCTURES OF RIBOSOMAL 5S RNA AND 5.8S RNA

核糖体 5S RNA 和 5.8S RNA 的结构

• 批准号: 3276838
• 负责人: ALAN G MARSHALL
• 金额: $ 10.64万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-01-01 至 1988-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3276838
• 关键词: Bacillus subtilis; Escherichia coli; Neurospora; Raman spectrometry; Saccharomyces; Trichomonas; circular dichroism; electron spin resonance spectroscopy; high performance liquid chromatography; infrared spectrometry; intermolecular interaction; light scattering; magnesium; mathematical model; microcalorimetry; nuclear magnetic resonance spectroscopy; nucleic acid sequence; nucleic acid structure; nucleobase; ribosomal RNA; thermodynamics; ultraviolet spectrometry; wheat; yeasts

Objectives: A concerted series of site-selective physical measurements on ribosomal RNA's grown and isolated from both eukaryotic and prokaryotic organisms is designed to identify specific secondary and tertiary architectural features of 5S RNA and 5.8S RNA in solution. The results should make it possible to establish a secondary base-pairing pattern general to all known 5S RNA and 5.8S RNA primary nucleotide sequences. This universal secondary structure could then form a basis for tracing phylogenetic pathways over an extremely long evolutionary time scale, since the presence and function of 5S RNA is so highly conserved for all organisms from which it has been isolated. Methods: Optical techniques (ultraviolet, Fourier transform infrared and Raman spectroscopy) give the approximate total number of base pairs and the relative and absolute numbers of GC and AU pairs. Proton NMR at 500 MHz gives the specific number, types and sequence (e.g., AU followed by GC) of base pairs (including GU pairs) from intra-and inter-pair homonuclear Overhauser enhancements. Stepwise unfolding of the molecule with temperature is monitored by 1H and 31p NMR, FT-IR, ESR and differential scanning microcalorimetry. ESR spin labeling at the 5S RNA terminus, and at specific residues aids in assignment of NMR signals and in determination of local flexibility at those sites. Nucleotide bases exposed at the RNA surface are detected by effects of paramagnetic agents or chemically induced dynamic nuclear polarization (CIDNP) effects on 1H NMR signals. Efforts to crystaliize several 5S RNA's (for eventual X-ray diffraction studies) are in progress. Thermodynamic stabilities will be computed theoretically for all 80-odd primary sequences, for each of several proposed base-pairing schemes. Finally, all methods are applied to 5S RNA from several species (e.g., B. subtilis, E. coli, Neurospora crassa, wheat germ, and yeasts (Saccharomyces carlsbergensis; Torulopsis utilis).

目标：一系列一系列现场选择性的物理测量 核糖体RNA的生长并与真核和原核生物分离 生物旨在识别特定的二级和第三级 溶液中5S RNA和5.8S RNA的建筑特征。 结果 应该使建立二级碱基配对模式成为可能 一般到所有已知的5S RNA和5.8S RNA原发性核苷酸序列。 然后，这种通用的二级结构可以构成追踪的基础 系统发育途径超过非常长的进化时间尺度，因为 5S RNA的存在和功能对于所有人都如此高度保守 与之隔离的生物体。 方法：光学技术（紫外线，傅立叶变换红外和 拉曼光谱法）给出了大约基本对的总数和 GC和AU对的相对和绝对数量。 质子NMR为500 MHz 给出特定的数字，类型和顺序（例如，Au，然后是GC） 碱基对基室内和配对之间的碱基对（包括GU对） 大刺的增强功能。 分子逐步展开 温度由1H和31p NMR，FT-IR，ESR和差异监测 扫描微钙化。 5S RNA末端的ESR自旋标记，并且 在特定残留物中有助于分配NMR信号和确定 这些站点的局部灵活性。 暴露于RNA的核苷酸碱基 通过顺磁剂或化学作用检测表面 诱导动态核极化（CIDNP）对1H NMR信号的影响。 努力使几个5S RNA晶体（最终用于X射线衍射 研究正在进行中。 将计算热力学稳定性 理论上对于所有80多个主序列，几个序列 提出的基本配对方案。 最后，将所有方法应用于5s RNA 来自几种物种（例如B. uttilis，大肠杆菌，Neurospora crassa，小麦 细菌和酵母（saccharomyces carlsbergensis; torulopsis utilis）。