相关实验视频
Updated: Jul 31, 2026

11:22
Measuring the Kinetics of mRNA Transcription in Single Living Cells
Published on: August 25, 2011
通过大肠杆菌RNA聚合酶对转录暂停和停止的单分子研究
R J Davenport1, G J Wuite, R Landick
1Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.
概括
单分子研究表明,RNA聚合酶表现出不同的转录速率和暂停行为. 可逆暂停作为一个关键的中间状态,影响细菌和真核生物的基因调节.
科学领域:
- 分子生物学分子生物学
- 生物物理学的生物物理.
- 遗传学 遗传学是一种遗传学.
背景情况:
- 基因调控依赖于RNA聚合酶对转录的精确控制.
- 了解单个RNA聚合酶分子的动态行为对于阐明转录机制至关重要.
研究的目的:
- 在转录过程中调查单个RNA聚合酶分子的实时动态.
- 描述单个RNA聚合酶分子的内在转录速率,暂停倾向和停止行为.
- 探索暂停作为转录中的动力中间体的作用.
主要方法:
- 使用先进的光学陷/流量控制视频显微镜技术.
- 实时监测Escherichia coliRNA聚合酶单个分子在延长模板长度上的转录.
主要成果:
- 在单个RNA聚合酶分子之间的内在转录率中显示出显著的异质性.
- 观察到不同RNA聚合酶分子中暂停和终止的不同倾向.
- 确定可逆暂停是活跃延长和转录停止之间的关键动态中间体.
- 在转录过程中揭示了RNA聚合酶的形态转移稳定性.
结论:
- 单分子转录研究为RNA聚合酶行为的异质性提供了关键的见解.
- RNA聚合酶的结构灵活性和暂停动力学是基因调节的基础.
- 这些发现对理解原生生物和真核生物系统中的基因调节机制有直接影响.
相关概念视频
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...
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...
Transcription Elongation Factors
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 into a...
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 into a...
Transcription Attenuation in Prokaryotes
Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure. Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
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...

