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High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
ドロソフィラ TAFII40は,VP16の活性化ドメインと基礎転写因子TFIIBの両方と相互作用する
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
まとめ
この研究では,TFIID複合体のサブユニットであるDrosophila TAFII40 (dTAFII40) と,そのトランスクリプションにおける役割が特定されています. dTAFII40は,ウイルス活性化剤VP16と基礎因子TFIIBを直接結合し,遺伝子調節に不可欠な三元相互作用を示唆しています.
科学分野:
- 分子生物学は分子生物学である.
- バイオケミストリー バイオケミストリー
- 遺伝学 遺伝学とは
背景:
- RNAポリメラーゼIIの転写は,様々なタンパク質複合体によって調節されます.
- TATA結合タンパク質 (TBP) とTBP関連因子 (TAFs) を含むTFIID複合体は,転写開始に不可欠である.
- VP16のようなウイルスのトランスアクティベーターは,トランスクリプション機構の構成要素と相互作用することによって,トランスクリプションを強化します.
研究 の 目的:
- TFIID複合体のサブユニットであるDrosophila TAFII40 (dTAFII40) をクローンし,発現し,生化学的に特徴づけること.
- dTAFII40のウイルス活性化剤および基礎転写因子とのタンパク質相互作用を調査する.
- アクティベータ媒介による転写強化におけるdTAFII40の役割を明らかにする.
主な方法:
- ドロソフィラ TAFII40.40.の分子クローニングと発現
- インビトロタンパク質-タンパク質相互作用アッセイ.
- アフィニティクロマトグラフィー. アフィニティクロマトグラフィー.
- 抗体阻害アッセイ 抗体阻害アッセイ
主要な成果:
- dTAFII40がVP16活性化ドメインに直接結合することが示されました.
- dTAFII40は基礎転写因子TFIIBに直接結合する.
- VP16,dTAFII40,TFIIBを含む三元相互作用が提案されました.
- dTAFII40に対する抗体は,GAL4-VP16媒介の活性化を阻害したが,基礎転写を阻害しなかった.
結論:
- dTAFII40は,ドロソフィラ TFIID複合体の機能的サブユニットである.
- TFIID内のTAFは,アクティベーターと基礎転写因子の両方と相互作用することができます.
- 異なる活性化剤は,異なるTAFと相互作用し,特定の転写調節に貢献する可能性があります.
関連する概念動画
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...

