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Updated: May 27, 2026

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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 σサブユニットによるプロモーター-10要素認識の構造的基礎
Andrey Feklistov1, Seth A Darst
1The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA. afeklistov@rockefeller.edu
Cell
|December 6, 2011
まとめ
バクテリアRNAポリメラーゼシグマサブユニットは,10のプロモーター要素を認識し,DNA開封を駆動する. 結晶構造は,A(-11) とT(-7) 塩基との重要な相互作用を明らかにし,転写の開始を説明しています.
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:
- バクテリアの転写開始は,遺伝子調節に不可欠です.
- RNAポリメラーゼシグマサブユニットの−10プロモーター要素への結合は,プロモーター開封に不可欠である.
- この相互作用の構造的基礎を理解することは,転写調節を解読する鍵です.
研究 の 目的:
- -10プロモーターDNAに結合したシグマサブユニットドメイン2の結晶構造を決定する.
- プロモーターの開きと転写の開始を駆動する分子相互作用を解明する.
主な方法:
- 高解像度構造を得るためのX線結晶学.
- 構造的な発見を検証するための生化学的分析.
主要な成果:
- 結晶構造は,シグマ領域2とDNAの脊髄の間の広範な相互作用を明らかにします.
- A(-11) とT(-7) との特定の塩基相互作用が特定され,これらの塩基は,タンパク質ポケットに転がり,埋められた.
- シグマによる−10要素認識が直接プロモーター開封を誘導するモデルが提案されています.
結論:
- この研究は,バクテリアのプロモーター溶解の詳細な構造的メカニズムを提供します.
- A(-11) とT(-7) は,配列認識とDNA挤出において重要な役割を果たしています.
- これらの発見は,開始時に転写バブルの形成に関する重要な洞察を提供します.
関連する概念動画
The Eukaryotic Promoter Region
The eukaryotic promoter region is a segment of DNA located upstream of a gene. It contains an RNA polymerase binding site, a transcription start site, and several cis-regulatory sequences. The proximal promoter region is located in the vicinity of the gene and has cis-regulatory sequences and the core promoter. The core promoter is the binding site for RNA polymerase and is usually located between -35 and +35 nucleotides from the transcription start site. The distal promoter regions are...
The Eukaryotic Promoter Region
The eukaryotic promoter region is a segment of DNA located upstream of a gene. It contains an RNA polymerase binding site, a transcription start site, and several cis-regulatory sequences. The proximal promoter region is located in the vicinity of the gene and has cis-regulatory sequences and the core promoter. The core promoter is the binding site for RNA polymerase and is usually located between -35 and +35 nucleotides from the transcription start site. The distal promoter regions are...
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:
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...

