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转录的结构基础:RNA聚合酶II在2.8安格斯特罗姆分辨率下
P Cramer1, D A Bushnell, R D Kornberg
1Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305-5126, USA.
概括
酵母RNA聚合酶II的结构研究揭示了四个移动模块,包括一个对转录启动至关重要的. 详细介绍了活性部位的金属离子和RNA退出通路.
科学领域:
- 分子生物学分子生物学
- 结构生物学 结构生物学
- 生物化学 生物化学
背景情况:
- RNA聚合酶II对于真核生物的基因转录至关重要.
- 了解它的结构是解读转录过程的关键.
- 之前的研究表明,RNA聚合酶II中存在移动管域.
研究的目的:
- 为了确定酵母RNA聚合酶II的高分辨率结构.
- 阐明转录启动和延长的结构基础.
- 在酶复合体内识别关键的功能元素.
主要方法:
- 使用X射线晶体学,以2.8和3.1安格斯特罗姆分辨率获得结构.
- 对不同晶体形式的比较分析揭示了移动模块.
- 区别 富里埃映射在活性位点确定了金属离子.
主要成果:
- 酵母RNA聚合酶II结构包括四个移动模块,包括一个摇摆.
- 在2.8安格斯特罗姆结构中的一个开放的形结构可以促进促进者DNA的进入.
- 在活性部位观察到两种金属离子,其中一种在RNA合成过程中可能可交换.
- 有证据表明,RNA退出发生在碳素终端重复域附近,将合成与处理联系起来.
结论:
- 已识别的移动模块,特别是,在转录中起着至关重要的作用.
- 这些结构性见解为理解DNA/RNA相互作用和金属离子催化提供了基础.
- 这些发现表明RNA合成和RNA处理机械之间的功能合.
相关概念视频
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...
RNA Polymerase II Accessory Proteins
Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
Bacterial Transcription
RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:
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...
RNA Polymerase II Accessory Proteins
Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...

