関連する実験動画
Updated: Jul 15, 2026

07:44
High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
RAP74をコードするcDNAが,RNAポリメラーゼIIによる転写の一般的始動因子である
A Finkelstein1, C F Kostrub, J Li
1Department of Biochemistry, Michigan State University, E. Lansing 48824.
Nature
|January 30, 1992
まとめ
研究者は,トランスクリプション因子RAP30/74 (TFIIF) の重要な構成要素であるヒトRAP74のcDNAを分離しました. この因子は,RNAポリメラーゼII前始動複合体の形成と正確な転写開始に不可欠です.
科学分野:
- 分子生物学は分子生物学である.
- 遺伝子規制 遺伝子規制
- バイオケミストリー バイオケミストリー
背景:
- RNAポリメラーゼIIは,一般の転写因子を初期化するために必要とします.
- RAP30/74 (TFIIF) は,プレイニシアチブ複合体の組み立てに不可欠な一般的な転写因子です.
- RAP30サブユニットは,バクテリアのシグマ因子とホモロジーを共有しています.
研究 の 目的:
- 人間のRAP74サブユニットをコードする補完DNA (cDNA) を分離する.
- トランスクリプション開始におけるRAP30とRAP74の役割を特徴づける.
- トランスクリプションアッセイにおける再結合RAP30とRAP74の有用性を評価する.
主な方法:
- 補完的なDNA (cDNA) の分離と配列決定.
- エシェリキヤ大腸における再結合タンパク質発現.
- RNAポリメラーゼIIを用いたインビトロ転写アッセイ.
主要な成果:
- 人間のRAP74cDNAを分離した.
- RAP30とRAP74の両方がプレイニシアチブ・コンプレックスの不可欠な構成要素であることを実証する.
- 再結合ヒトRAP30とRAP74は,トランスクリプション開始における自然複合体の機能を機能的に置き換えることができる.
結論:
- 人間のRAP74cDNAの分離は,TFIIF.のさらなる研究のためのツールを提供します.
- 再結合TFIIFはインビトロで機能し,詳細なメカニズム研究を可能にします.
- TFIIFの機能を理解することは,RNAポリメラーゼIIによる遺伝子転写の調節を解読する上で極めて重要です.
関連する概念動画
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...
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...
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...
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...

