相关实验视频
Updated: Jul 26, 2026

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High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
通过RNA聚合酶II启动转录的五个中间复合体
S Buratowski1, S Hahn, L Guarente
1Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.
Cell
|February 24, 1989
概括
研究人员在腺病毒转录启动过程中确定了五个顺序复合体. 这些复合体涉及一般转录因子和RNA聚合酶II,揭示了因子组合的顺序和转录启动的拟议模型.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 遗传学 是一个遗传学.
背景情况:
- 转录启动是一个复杂的过程,涉及多种蛋白质因素.
- 了解这些因素的精确组装顺序对于破译基因调节至关重要.
研究的目的:
- 解决和描述在腺病毒主要晚期促销者的转录启动复合体的顺序形成.
- 确定这些复合体内的一般转录因子和RNA聚合酶II的相对位置.
主要方法:
- 使用原生凝电泳DNA结合试验来识别不同的复杂物.
- 用DNAase I足迹分析来绘制结合点和复合体内的因子的相对位置.
主要成果:
- 确定了五个不同的复合体,由转录因子 (TFIID,TFIIA,TFIIB,RNA聚合酶II,TFIIE) 的顺序添加形成.
- TFIIA 和 TFIID 与 TATA 元素结合,TFIIB 作为与 RNA 聚合酶 II 的桥梁.
- ATP/dATP和TFIIE结合改变了DNAse I足迹,并添加了核糖核酸三酸盐,导致形成具有启动转录的复合体.
结论:
- 提出了一种用于组装转录前启动复合物的序列模型.
- 该研究阐明了在腺病毒转录启动过程中一般转录因子和RNA聚合酶II的动态相互作用.
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...
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...
General Transcription Factors
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
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...

