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Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
通过单个亚单元RNA聚合酶启动转录的反应途径上的时间解析事件:拉曼晶体学证据
Yuanyuan Chen1, Ritwika Basu, Michael L Gleghorn
1Department of Biochemistry, Case Western Reserve University, Cleveland, Ohio 44106, United States.
Journal of the American Chemical Society
|July 13, 2011
概括
这项研究使用拉曼显微镜观察N4菌体RNA聚合酶 (RNAP) 中的实时核类转移反应. 这些发现揭示了基键形成的关键步骤和催化过程中的酶构造变化.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 结构生物学 结构生物学
背景情况:
- N4菌体RNA聚合酶 (RNAP) 对于病毒RNA复制至关重要.
- 了解基结形成的动力学是理解RNAP功能的关键.
研究的目的:
- 为了实时监测N4菌体RNAP单晶体中的核基转移反应.
- 为了阐明在催化过程中的形状变化和基质/金属结合动态.
主要方法:
- 单个RNAP-DNA复合晶体的实时拉曼显微镜.
- 在晶体中浸泡核三酸盐 (NTP) 基质和双价离子.
- 利用同位素标记的NTP和X射线晶体学数据进行光谱分配.
主要成果:
- 在2-7分钟内观察到基质结合和O螺旋形状变化.
- 追踪双价金属结合和三酸糖酸盐形状的变化.
- 二键的形成在15分钟内完成,金属在40分钟后释放.
结论:
- 只有通过基质结合才能诱导O螺旋运动.
- 晶体内反应动力学揭示了暂时的中间体,以便进一步研究.
- 这项工作提供了RNAP催化在分子水平的动态视图.
相关概念视频
Transcription Initiation
Initiation is the first step of transcription in eukaryotes. Prokaryotic RNA Polymerase (RNAP) can bind to the template DNA and start transcribing. On the other hand, transcription in eukaryotes requires additional proteins, called transcription factors, to first bind to the promoter region in the DNA template. This binding helps recruit the specific RNAP that can assemble on the DNA and start transcription.
The promoters and enhancers and their accessory proteins allow tight regulation of...
The promoters and enhancers and their accessory proteins allow tight regulation of...
Eukaryotic RNA Polymerases
RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
All three eukaryotic RNAPs require specific transcription factors, of which the...
Eukaryotic RNA Polymerases
RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
All three eukaryotic RNAPs require specific transcription factors, of which the...
Bacterial RNA Polymerase
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Bacterial RNA Polymerase
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
Ribosomal RNA Synthesis
Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...

