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Updated: May 9, 2026

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High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
一种哺乳动物RNA聚合酶II全酶,含有促进体特异转录启动所需的所有成分
V Ossipow1, J P Tassan, E A Nigg
1Département de Biologie Moléculaire, Sciences II, Université de Genéve, Switzerland.
Cell
|October 6, 1995
概括
研究人员在哺乳动物细胞中确定了一个完整的RNA聚合酶II全酶,这表明真核生物和 prokaryotic 转录启动之间的相似之处. 这个复合体包括对基因表达至关重要的关键转录因子.
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:
- 蛋白激酶MO15/CDK7与转录因子TFIIH有关.
- MO15/CDK7可酸化RNA聚合酶II的碳氧终端域.
研究的目的:
- 调查MO15/CDK7与RNA聚合酶II转录机制的关联.
- 在哺乳动物细胞中识别RNA聚合酶II全酶的成分.
主要方法:
- 使用抗MO15/CDK7抗体进行单克隆抗体共免疫沉降.
- 对老鼠肝核提取物的分析.
主要成果:
- MO15/CDK7抗体共免疫降落RNA聚合酶II,TFIID,TFIIB,TFIIH,TFIIF,和TFIIE. 这几种抗体的组分为:
- 免疫沉积物缺乏转录激活蛋白质,如HNF1,HNF4和C/EBPαα.
- 发现了哺乳动物细胞中自主启动RNA聚合酶II全酶的证据.
结论:
- 哺乳动物细胞含有一种能够自主启动的RNA聚合酶II全酶.
- 这一发现表明真核细胞和 prokaryotic 转录启动之间的功能平行.
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...

