CHARACTERIZATION OF LIGAND RECOGNITION OF A METABOLITE-RESPONSIVE RIBOZYME

代谢物响应核酶的配体识别特征

• 批准号: 7725203
• 负责人: JEFF SOUKUP
• 金额: $ 3.94万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7725203
• 关键词: 5' Untranslated Regions; Address; Affect; Amines; Catalysis; Catalytic RNA; Cell Wall; Cells; Computer Retrieval of Information on Scientific Projects Database; Enzymes; Funding; Gene Expression; Genetic; Glucosamine; Glucose; Gram-Positive Bacteria; Grant; Institution; Length; Ligands; Measures; Mediating; Messenger RNA; Metabolic; Molecular; Names; Proteins; RNA; Rate; Reaction; Research; Research Personnel; Resources; Role; Source; Translational Repression; United States National Institutes of Health; base; functional group; glucosamine 6-phosphate; inorganic phosphate; interest; novel; response; serinol

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The metabolic state of a cell typically affects gene expression through protein-mediated mechanisms of transcriptional or translational control. Yet recent studies have shown that messenger RNAs (mRNAs) can directly sense and respond to specific metabolites through intrinsic domains termed riboswitches. In the presence of metabolites, structural changes in riboswitch domains can result in either transcriptional termination or translational repression. Recently a novel catalytic riboswitch has been discovered that exerts genetic control through self-cleavage of the nascent RNA in response to cellular metabolite concentration. The metabolite-dependent ribozyme resides in the 5'-untranslated region of the glmS mRNA of numerous Gram-positive bacteria and it catalyzes an internal phosphoester transfer reaction that results in cleavage and inactivation of the mRNA. The ribozyme selectively recognizes and is 1000-fold activated by glucosamine-6-phosphate, the metabolic product of the GlmS enzyme, and an important component of bacterial cell walls. We are interested in understanding the molecular basis of ligand recognition and catalysis by the glmS riboswitch. To address this we measured the rate of self-cleavage of the ribozyme/riboswitch in response to a panel of related but distinct ligands, serinol, glucose and glucosamine, to name a few. We have determined that the ribozyme makes important contacts to the amine and phosphate in the natural ligand. We are interested in determining how these two functional groups are interacting with the full-length glmS ribozyme in order to fully understand its role in catalysis.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 细胞的代谢状态通常通过转录或翻译控制的蛋白质介导的机制影响基因表达。 然而，最近的研究表明，Messenger RNA（mRNA）可以通过称为核糖开关的内在域直接感知并响应特定的代谢产物。 在存在代谢物的情况下，核糖开关结构域的结构变化可能导致转录终止或翻译抑制。 最近，已经发现一种新型的催化核糖开关，该核糖开关通过对细胞代谢产物浓度的新生RNA的自切除来发挥遗传控制。 代谢产物依赖性核酶属于许多革兰氏阳性细菌的GLMS mRNA的5'-非翻译区域，并且它催化了内部磷酸酯转移反应，从而导致mRNA的切割和失活。 核酶有选择地识别，并被葡萄糖胺-6-磷酸盐，GLMS酶的代谢产物和细菌细胞壁的重要组成部分激活1000倍。 我们有兴趣了解GLMS核糖开关的配体识别和催化的分子基础。 为了解决这个问题，我们测量了核酶/核糖开关的自裂速率，以响应一组相关但不同的配体，塞氨酸，葡萄糖和葡萄糖胺，仅举几例。 我们已经确定核酶在天然配体中与胺和磷酸盐之间的重要接触。 我们有兴趣确定这两个官能团如何与全长GLMS核酶相互作用，以充分了解其在催化中的作用。