関連する実験動画
Updated: Oct 2, 2026

07:44
High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
バクテリアのRNAポリメラーゼホロ酵素とRNAポリメラーゼプロモーターの構造的組織 オープンコンプレックス
Vladimir Mekler1, Ekaterine Kortkhonjia, Jayanta Mukhopadhyay
1Howard Hughes Medical Institute, Waksman Institute, Rutgers University, Piscataway, NJ 08854, USA.
Cell
|March 15, 2002
まとめ
研究者は,バクテリアのRNAポリメラーゼホロ酵素とそのプロモーターオープン複合体の3D構造をマッピングしました. キーシグマ70) 領域1.1は劇的にシフトし,DNA結合における役割を明らかにする.
科学分野:
- 分子生物学は分子生物学である.
- 構造生物学 構造生物学とは
- バイオケミストリー バイオケミストリー
背景:
- 細菌のRNAポリメラーゼホロ酵素は,転写の開始に不可欠です.
- シグマ (70) 相互作用の構造的ダイナミクスを理解することは,転写調節を解読する鍵です.
研究 の 目的:
- バクテリアのRNAポリメラーゼホロ酵素とRNAポリメラーゼプロモーターの3次元構造を,溶液中のオープンコンプレックスで決定する.
- 核酵素とDNAとの相互作用の構造的基礎を解明する.
主な方法:
- 体系的な光共振エネルギー転送 (FRET) が採用されました.
- 距離制限ドッキングは,構造をモデル化するために使用されました.
主要な成果:
- ホロ酵素とオープン複合体の3次元構造が定義されました.
- シグマ70) の1.2,2,3,4の領域は,両州で類似したポジションを示した.
- シグマ70) 領域1.1は,ホロ酵素の活性センター裂け目と,オープン複合体の外側に位置する,重要な位置差を示した.
結論:
- この研究は,シグマ (70) 相互作用を理解するための構造的枠組みを提供します.
- シグマ70) 領域1.1の独特な位置付けは,DNA結合と転写の開始における規制的役割を示唆する.
- この方法論は,他の大型分子複合体の研究にも適用できます.
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...
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...
Bacterial Transcription
RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:
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...
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...
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...

