古代RNA聚合酶转录延长和Spt4/5招募的结构基础
相关实验视频
Updated: Jun 27, 2025

07:44
High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
14.2K
Daniela Tarău1, Felix Grünberger1, Michael Pilsl2
1Institute of Microbiology & Archaea Centre, Single-Molecule Biochemistry Lab, University of Regensburg, 93053 Regensburg, Germany.
Nucleic acids research
|May 6, 2024
概括
考古转录涉及RNA聚合酶 (RNAP) 和Spt4/5.5的过程. 结构分析显示,即使没有核酸,Spt4/5也与RNAP结合,影响转录泡的稳定性.
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:
- 考古转录利用RNA聚合酶 (RNAP),同类于真核RNAP II.
- 在所有生命领域中存在的Spt5,在古生物和真核生物中与Spt4异构.
- 了解古生物中的Spt4/5功能对于破译转录机制至关重要.
研究的目的:
- 阐明 Spt4/5 函数在考古转录中的机械基础.
- 为Spt4/5与古代RNAP.的相互作用提供结构性见解.
主要方法:
- 单粒子冷电子显微镜 (cryo-EM) 的重建.
- 使用Pyrococcus furiosus (Pfu) 转录机器进行结构功能分析.
- 在体外生化测试.
主要成果:
- Pfu Spt4/5以超收缩的形状与ApoRNAP结合,独立于核酸.
- RNAP表现出形状灵活性,在EC形成或Spt4/5结合时收缩.
- Spt5-NGN域与DNA接触,稳定转录泡并影响Spt4/5活动.
结论:
- 在考古转录中Spt4/5功能的结构基础被阐明.
- Spt4/5 在稳定转录泡方面发挥作用,可能超出延长支.
- RNAP中的 conformational 变化是Spt4/5介导的转录调节的关键.
相关概念视频
Transcription Initiation
16.4K
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...
16.4K
Eukaryotic RNA Polymerases
24.1K
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...
24.1K
Bacterial RNA Polymerase
29.5K
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...
29.5K
Transcription Elongation Factors
10.8K
Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA...
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA...
10.8K
Bacterial Transcription
28.1K
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:
28.1K
RNA Polymerase II Accessory Proteins
9.2K
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...
9.2K

