DEFINING THE ROLE PLAYED BY 23S RRNA IN TRANSLATION
定义 23S RRNA 在翻译中所扮演的角色
基本信息
- 批准号:6182196
- 负责人:
- 金额:$ 21.3万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1999
- 资助国家:美国
- 起止时间:1999-05-01 至 2004-04-30
- 项目状态:已结题
- 来源:
- 关键词:Bacillus stearothermophilus active sites aminoacyl tRNA aminoacyltransferase chemical kinetics enzyme mechanism enzyme substrate enzyme substrate complex genetic translation nucleic acid reconstitution nucleic acid sequence nucleic acid structure ribosomal RNA ribosomes site directed mutagenesis transfer RNA
项目摘要
Translation of the genetic information in the cell into functional proteins is an essential and highly conserved function. The ribosome, a two-subunit macromolecular complex, composed in bacteria of three large RNAs (rRNAs) and more than 50 proteins (r-proteins), is the catalyst and framework for the intricate and coordinated process of translation. Peptidyl transferase, likely the most primitive activity of the ribosome, is of central importance. Theoretical considerations of the origin of life predict a central role for the rRNAs in translation, a prediction strengthened by the demonstration of catalysis by RNA and consistent with the extreme phylogenetic conservation observed among rRNA sequences. Extensive biochemical and genetic studies indicate that the rRNAs play a primary functional role in the processes of translation, and in particular 23S rRNA in the central catalytic event, peptide bond formation. Structural studies of the ribosome, including cryoelectron microscopy and X-ray crystallography, are yielding a wealth of information. However, the static views of the ribosome yielded by such approaches will not reveal the mechanism or dynamics of the intricate process of translation. This proposal describes in vitro genetic and biochemical approaches to identify the molecular components at the active site of peptidyl transferase (RNA or protein). The goal of these studies is to understand how these components conspire in the formation of peptide bonds and to understand how this activity is controlled between each catalytic cycle to allow for the generation of high-fidelity encoded protein products. First, site-directed mutagenesis and modification interference approaches will be used to identify nucleotides in 23S rRNA that are specifically involved in tRNA substrate binding and catalysis by the ribosome. Second, a recently developed iterative in vitro selection approach will be used to optimize the activity of ribosomes reconstituted from in vitro transcribed Bacillus stearothermophilus 23S rRNA. Finally, a variety of biochemical approaches including crosslinking, kinetic studies and chemical modification will be used to characterize the structure and function of the hybrid state of tRNA binding in translational elongation. Information obtained from these studies will eventually help to define the features of current ribosome- specific antibiotics which make them effective and the mechanism by which these same antibiotics are evaded by the host. Ultimately, this will lead to the rational design of novel antibiotics.
将细胞中的遗传信息转化为功能蛋白是一项重要且高度保守的功能。 核糖体是一种二亚基大分子复合物,由细菌中的三种大RNA (rRNA) 和超过50 种蛋白质(r-蛋白质) 组成,是复杂且协调的翻译过程的催化剂和框架。 肽基转移酶可能是核糖体最原始的活性,具有至关重要的作用。 生命起源的理论考虑预测了 rRNA 在翻译中的核心作用,这一预测因 RNA 催化作用的证明而得到加强,并且与在 rRNA 序列中观察到的极端系统发育保守性一致。 广泛的生化和遗传学研究表明,rRNA 在翻译过程中发挥着主要功能作用,特别是 23S rRNA 在核心催化事件(肽键形成)中。 核糖体的结构研究,包括冷冻电子显微镜和 X 射线晶体学,正在产生大量信息。 然而,通过这种方法产生的核糖体的静态视图不会揭示复杂的翻译过程的机制或动态。该提案描述了体外遗传和生化方法来鉴定肽基转移酶(RNA 或蛋白质)活性位点的分子成分。 这些研究的目的是了解这些成分如何共同形成肽键,并了解如何在每个催化循环之间控制这种活性,以生成高保真编码的蛋白质产物。 首先,定点诱变和修饰干扰方法将用于鉴定 23S rRNA 中专门参与 tRNA 底物结合和核糖体催化的核苷酸。 其次,最近开发的迭代体外选择方法将用于优化由体外转录的嗜热脂肪芽孢杆菌 23S rRNA 重组的核糖体的活性。 最后,包括交联、动力学研究和化学修饰在内的多种生化方法将用于表征翻译延伸中 tRNA 结合的杂合状态的结构和功能。 从这些研究中获得的信息最终将有助于确定当前核糖体特异性抗生素的特征(使其有效)以及宿主逃避这些抗生素的机制。最终,这将导致新型抗生素的合理设计。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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RACHEL GREEN其他文献
RACHEL GREEN的其他文献
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{{ truncateString('RACHEL GREEN', 18)}}的其他基金
Biochemistry, Cellular and Molecular Biology Program: JHU BioGREAT (Biomedical Graduate REsiliency & Adaptability Training)
生物化学、细胞和分子生物学项目:JHU BioGREAT(生物医学研究生 REsiliency
- 批准号:
10810143 - 财政年份:2022
- 资助金额:
$ 21.3万 - 项目类别:
Biochemistry, Cellular and Molecular Biology Program
生物化学、细胞和分子生物学项目
- 批准号:
10332103 - 财政年份:2022
- 资助金额:
$ 21.3万 - 项目类别:
Biochemistry, Cellular and Molecular Biology Program
生物化学、细胞和分子生物学项目
- 批准号:
10650714 - 财政年份:2022
- 资助金额:
$ 21.3万 - 项目类别:
Kinetic Dissection of Eukaryotic Translation Initiation
真核翻译起始的动力学剖析
- 批准号:
8469510 - 财政年份:2000
- 资助金额:
$ 21.3万 - 项目类别:
DEFINING THE ROLE PLAYED BY 23S RRNA IN TRANSLATION
定义 23S RRNA 在翻译中所扮演的角色
- 批准号:
2835607 - 财政年份:1999
- 资助金额:
$ 21.3万 - 项目类别:
The role played by the RNAs and tRNAs in translation
RNA 和 tRNA 在翻译中发挥的作用
- 批准号:
7529208 - 财政年份:1999
- 资助金额:
$ 21.3万 - 项目类别:
The role played by the RNAs and tRNAs in translation
RNA 和 tRNA 在翻译中发挥的作用
- 批准号:
7031934 - 财政年份:1999
- 资助金额:
$ 21.3万 - 项目类别:
The role played by the RNAs and tRNAs in translation
RNA 和 tRNA 在翻译中发挥的作用
- 批准号:
7151944 - 财政年份:1999
- 资助金额:
$ 21.3万 - 项目类别:
The role played by the RNAs and tRNAs in translation
RNA 和 tRNA 在翻译中发挥的作用
- 批准号:
8010950 - 财政年份:1999
- 资助金额:
$ 21.3万 - 项目类别:
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