関連する実験動画
Updated: May 17, 2026

07:44
High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
転写開始の構造的基礎
Yu Zhang1, Yu Feng, Sujoy Chatterjee
1Howard Hughes Medical Institute, Waksman Institute, and Department of Chemistry and Chemical Biology, Rutgers University, Piscataway, NJ 08854, USA.
まとめ
この研究では,RNAポリメラーゼ (RNAP) 転写開始複合体の結晶構造が明らかになりました. これらの構造は,RNAPとシグマ因子 (σ) が,転写を開始するためのプロモーターDNA要素を認識する方法を詳細に説明しています.
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:
- トランスクリプションの開始は,遺伝子発現における重要な規制段階です.
- RNAポリメラーゼ (RNAP) は,転写を開始するためにプロモーターDNAとオープンコンプレックスを形成します.
- このプロセスの構造的基礎を理解することは,遺伝子調節を解読する鍵です.
研究 の 目的:
- 機能的転写開始複合体の高解像度結晶構造を決定する.
- RNAP,シグマ因子 (σ),およびプロモーターDNAの間の分子相互作用を解明する.
- プロモーター認識とDNA解のメカニズムについての洞察を提供するため.
主な方法:
- 構造を特定するために,X線結晶学を用いた.
- Thermus thermophilus RNAポリメラーゼ,σ(A),およびプロモーターDNAの機能的転写始動複合体を利用した.
- 構造は2.9と3.0 Åの解像度で解像しました.
主要な成果:
- 結晶構造は,σが−10要素と差別要素との特殊な相互作用を明らかにし,DNA塩基の解離とポケット挿入を含む.
- RNAPと -4/+2領域の相互作用が特定され, +2塩基がRNAPポケットに挿入された.
- σとテンプレート鎖単一鎖DNA (ssDNA) の間の相互作用は,RNAP活性センターのためのssDNAを前編成することが示されました.
結論:
- 決定された構造は,転写開始時にプロモーター認識の詳細な分子視点を提供します.
- これらの発見は,プロモーターDNAの結合と解き放たれにおける σ と RNAP の正確な役割を強調しています.
- この研究は,トランスクリプション開始複合体が効率的なDNAエンゲージメントのためにどのように事前に組織されているかを理解するための構造的基礎を提供します.
関連する概念動画
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...
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:
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...
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...
Eukaryotic Transcription Activators
Transcription activators are proteins that promote the transcription of genes from DNA to RNA. In most cases, these proteins contain two separate domains ‒ a domain that binds to DNA and a domain for activating transcription; however, in some cases, a single domain is responsible for both binding and activation of transcription, as seen in the glucocorticoid receptor and MyoD.
The binding domains are capable of recognizing and interacting with regulatory sequences on the DNA. These domains are...
The binding domains are capable of recognizing and interacting with regulatory sequences on the DNA. These domains are...

