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Substrate Recognition of a tRNA Modifying Enzyme

tRNA 修饰酶的底物识别

基本信息

批准号：
7104456
负责人：
George A Garcia
金额：
$ 27.19万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-08-01 至 2008-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7104456
关键词：
biochemical evolution crystallization enzyme mechanism enzyme structure enzyme substrate functional /structural genomics guanine analog isozymes pentosyltransferase proteomics radiotracer transfer RNA

项目摘要

DESCRIPTION (provided by applicant): Queuine (one of ca. 100 naturally occurring modified bases) is found in the wobble position (34) of 4 tRNAs (anticodons GUN) in eubacteria and eukaryotes. It is introduced into tRNAs by a post-transcriptional base exchange catalyzed by tRNA-guanine transglycosylase (TGT, also conserved across the kingdoms).. It has been shown that TGT is required for pathogenicity of the dysentery-causing bacterium Shigella flexneri. While the tgt gene has been shown not to be required for growth in cell culture, the relationship between tgt and pathogenicity is not understood, and the present data suggest that TGT could be a novel antibiotic target. Previous studies have elucidated many aspects of the molecular basis of the recognition and discrimination of specific tRNAs and heterocyclic substrates by the eubacterial TGT. However, much work still needs to be done to understand the differential properties of eukaryotic vs. eubacterial TGTs. Other than correlations with proliferation, differentiation, retroviral frameshifting, and bacterial virulence, little is known about the physiological roles of queuine let alone how these roles are carried out. There are two overarching goals of this proposal. The first is to elucidate the similarities and differences between the eubacterial TGT and the human TGT. These studies are prerequisite to the rational design of selective inhibitors of the eubacterial TGT. Our second overarching goal is to try to elucidate the physiological role(s) of queuine in eubacteria and eukaryotes. Given the potential of TGT as a novel antibiotic target, it is important to understand the physiological role(s) of queuine and any differences between human and eubacteria. In order to achieve these goals, we have proposed a number of specific aims that bring many techniques and approaches (e.g., molecular biology, enzymology, crystallography, computational chemistry, synthetic chemistry, genomics, and proteomics) to bear on this problem. To carry out these studies in a rigorous fashion, we have established collaborations with key experts and take advantage of the resources available here at the University of Michigan. A clear understanding of the similarities and differences between the TGTs from eubacteria and human will be of significance for many reasons but most obviously for the future design of novel antibiotics.

描述（由申请人提供）：Queuine（约 100 个天然存在的修饰碱基之一）发现于真细菌和真核生物中 4 个 tRNA（反密码子 GUN）的摆动位置（34）中。它通过 tRNA-鸟嘌呤转糖基酶（TGT，在各个王国中也保守）催化的转录后碱基交换引入 tRNA。研究表明，TGT 是引起痢疾的福氏志贺氏菌的致病性所必需的。虽然 tgt 基因已被证明不是细胞培养物中生长所必需的，但 tgt 和致病性之间的关系尚不清楚，目前的数据表明 TGT 可能是一种新的抗生素靶点。先前的研究已经阐明了真细菌 TGT 识别和区分特定 tRNA 和杂环底物的分子基础的许多方面。然而，仍然需要做很多工作来了解真核 TGT 与真细菌 TGT 的差异特性。除了与增殖、分化、逆转录病毒移码和细菌毒力的相关性之外，人们对队列的生理作用知之甚少，更不用说这些作用是如何发挥的了。该提案有两个总体目标。首先是阐明真细菌TGT和人类TGT之间的异同。这些研究是合理设计真细菌 TGT 选择性抑制剂的先决条件。我们的第二个总体目标是尝试阐明奎因在真细菌和真核生物中的生理作用。鉴于 TGT 作为新型抗生素靶点的潜力，了解队列的生理作用以及人类和真细菌之间的差异非常重要。为了实现这些目标，我们提出了许多具体目标，利用许多技术和方法（例如分子生物学、酶学、晶体学、计算化学、合成化学、基因组学和蛋白质组学）来解决这个问题。为了以严格的方式开展这些研究，我们与主要专家建立了合作，并利用密歇根大学的可用资源。出于多种原因，清楚地了解真细菌和人类 TGT 之间的异同具有重要意义，但最明显的是对于未来新型抗生素的设计。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Glutamate versus glutamine exchange swaps substrate selectivity in tRNA-guanine transglycosylase: insight into the regulation of substrate selectivity by kinetic and crystallographic studies.

谷氨酸与谷氨酰胺交换交换 tRNA-鸟嘌呤转糖基酶中的底物选择性：通过动力学和晶体学研究深入了解底物选择性的调节。

DOI：
10.1016/j.jmb.2007.09.062
发表时间：
2007
期刊：
Journal of molecular biology
影响因子：
5.6
作者：
Tidten,Naomi;Stengl,Bernhard;Heine,Andreas;Garcia,GeorgeA;Klebe,Gerhard;Reuter,Klaus
通讯作者：
Reuter,Klaus

Differential heterocyclic substrate recognition by, and pteridine inhibition of E. coli and human tRNA-guanine transglycosylases.