トランスクリブするT7RNAポリメラーゼの構造は,イニシエーションから延長への移行段階にある
Kimberly J Durniak1, Scott Bailey, Thomas A Steitz
1Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06520-8114, USA.
まとめ
構造的研究により,T7細菌のDNA依存RNAポリメラーゼ (T7 RNAP) が,転写開始と延伸の間の移行をどのように行っているかが明らかになった. 中間結晶構造は,プロモーター結合ドメインの回転を示し,RNA合成に対応しながらDNA接触を維持します.
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:
- T7細菌のDNA依存RNAポリメラーゼ (T7RNAP) は,転写開始と延長の間に重要な構成変化を経験します.
- この移行の正確なメカニズム,特にアミノ端末ドメインを含むことは,まだ完全に理解されていません.
研究 の 目的:
- 転写開始から延長への移行中のT7RNAPの中間構造状態を明らかにする.
- T7 RNAP.の構成動態に関する高解像度構造的洞察を提供するためです.
主な方法:
- X線結晶学を用いて,T7 RNAP.の構造を決定した.
- プロモーターDNAと短いRNAトランスクリプト (7および8核酸) を含むT7RNAPの複合体を分析した.
主要な成果:
- 結晶構造は,転写中にT7 RNAPの中間状態を明らかにした.
- プロモーター結合ドメイン (PBD) と結合プロモーターは,8核酸RNAトランスクリプトの合成時に約45度回転する.
- この回転は,プロモーターの接触を維持し,成長するRNA-DNAヘテロデュプレックスの活性部位を拡張します.
結論:
- 観察された構造の変化は,T7 RNAPの経路を,開始から延長まで明らかにしています.
- PBDの回転は,本質的なDNAの相互作用を保ちながら,移行に対応するための重要なメカニズムです.
- サブドメインHは,イニシアチブのような位置にとどまり,アミノ端末ドメインの非均一な移行を示しています.
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...
Transcription Elongation Factors
Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
Transcription Elongation Factors
Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
The transcription elongation is regulated via pausing of RNA polymerase on several occasions during transcription. In bacteria, these halts are necessary because the transcription of DNA into mRNA is coupled to the translation of that mRNA into a...
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 Attenuation in Prokaryotes
Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure. Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
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...


