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High-throughput Purification of Affinity-tagged Recombinant Proteins
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通过西格玛区域1.2的非最佳结合来调节rRNA促进体:RNA聚合酶的额外识别元素
Shanil P Haugen1, Melanie B Berkmen, Wilma Ross
1Department of Bacteriology, University of Wisconsin-Madison, 420 Henry Mall, 53706, USA.
Cell
|June 17, 2006
概括
研究人员发现了大肠杆菌RNA聚合酶 (RNAP) 和细菌促进体之间的新相互作用. 这一发现揭示了促进体DNA序列如何影响RNAP结合,影响基因转录调节.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物化学 生物化学
背景情况:
- 转录启动调节通常涉及蛋白质结合DNA.
- 替代的特异性机制包括与RNA聚合酶 (RNAP) 结合的调节剂.
- 在RNAP促进体复合体中的动力变化提供了调节潜力.
研究的目的:
- 在细菌促进体中识别新型识别元素.
- 阐明特定序列RNAP相互作用的机制.
- 了解这种相互作用如何影响转录调节.
主要方法:
- 对rRNA促进体的突变发生.
- 对RNAP促进体复合物的动态分析.
- 二甲基硫酸盐足迹和交叉链接研究.
主要成果:
- 确定了1.2区域的西格玛子单元和rRNA促进体的区分区之间的新相互作用.
- 这种相互作用调节RNAP促进体复合物的稳定性.
- 不理想的相互作用导致对调节至关重要的短暂复合体.
结论:
- 西格玛子单位区域1.2作为促进者识别元素.
- 西格玛1.2分辨器相互作用微调RNAP促进体复合体的稳定性.
- 这种机制对于调节因细胞信号而产生的核糖体合成至关重要.
相关概念视频
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...
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 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...
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...
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...

