相关实验视频
Updated: May 10, 2026

07:44
High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
草TAFII40与VP16激活域和基底转录因子TFIIB相互作用
J A Goodrich1, T Hoey, C J Thut
1Howard Hughes Medical Institute, Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Cell
|November 5, 1993
概括
这项研究确定了Drosophila TAFII40 (dTAFII40),TFIID复合物的子单元,以及它在转录中的作用. dTAFII40直接与病毒激活剂VP16和基底因子TFIIB结合,这表明三元相互作用对基因调节至关重要.
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:
- RNA聚合酶II转录是由各种蛋白质复合体调节的.
- 由TATA结合蛋白 (TBP) 和TBP相关因子 (TAF) 组成的TFIID复合体对于转录启动至关重要.
- 像VP16这样的病毒转换激活剂通过与转录机制的组件相互作用来增强转录.
研究的目的:
- 克隆,表达,并生物化学地表征Drosophila TAFII40 (dTAFII40),这是TFIID复合体的一个子单元.
- 研究dTAFII40与病毒激活剂和基底转录因子的蛋白质-蛋白质相互作用.
- 阐明dTAFII40在激活剂介导的转录增强中的作用.
主要方法:
- 分子克隆和Drosophila TAFII40.40的表达
- 在体外蛋白质与蛋白质相互作用测试.
- 亲和性染色学. 亲和性染色学.
- 抗体抑制试验. 抗体抑制试验.
主要成果:
- 证明了dTAFII40与VP16激活域的直接结合.
- dTAFII40直接与基底转录因子TFIIB结合.
- 一个涉及VP16,dTAFII40和TFIIB的三元相互作用被提出.
- 对dTAFII40的抗体抑制了GAL4-VP16介导的激活,但不能抑制基底转录.
结论:
- dTAFII40 是 Drosophila TFIID 复合体的一个功能子单元.
- 在TFIID中的TAF可以与激活剂和基底转录因子相互作用.
- 不同的激活剂可能与不同的TAF相互作用,有助于特定的转录调节.
相关概念视频
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...
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...
Master Transcription Regulators
Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
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...
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...
Master Transcription Regulators
Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...

