在内源酵母基因上实时观察转录启动和延长
Daniel R Larson1, Daniel Zenklusen, Bin Wu
1Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
概括
研究人员开发了一种新的方法来观察酵母基因上的单RNA聚合酶活性. 他们发现基因转录启动没有记忆,延长速度有所不同,因子搜索时间控制了基因发射率.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物物理学的生物物理.
背景情况:
- 细胞信使RNA (mRNA) 水平是复杂的转录调节的结果.
- 小分子通路中的随机波动使理解基因表达动态变得复杂.
- 在体内观察内源性基因的单聚合酶活性在实验上具有挑战性.
研究的目的:
- 开发和应用一种方法来观察活跃酵母位上新生RNA的动态.
- 阐明转录启动,延长和终止的体内机制.
- 研究转录因子动态和基因表达率之间的关系.
主要方法:
- 光标记RNA的波动分析.
- 在内源性酵母基因中实时观察新生RNA动态.
- 测量上游转录因子丰度和核内移动性的测量.
主要成果:
- 这项研究成功地测量了新生RNA的启动,延长和终止动态.
- 在个别基因启动事件之间没有观察到转录记忆.
- 延长速度在整个细胞周期中显示出三倍的变化.
- 发现基因激活率与转激活因子搜索时间直接相关.
结论:
- 开发的方法允许实时观察内源基因单聚合酶活性.
- 酵母基因转录启动是一个独立的事件,没有记忆.
- 转录因子搜索动态是基因表达水平的关键决定因素.
相关概念视频
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...
Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...
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...
Yeast Signaling
Yeasts are single-celled organisms, but unlike bacteria, they are eukaryotes (cells with a nucleus). Cell signaling in yeast is similar to signaling in other eukaryotic cells. A ligand, such as a protein or a small molecule released from a yeast cell, attaches to a receptor on the cell surface. The binding stimulates second-messenger kinases to activate or inactivate transcription factors that further regulate gene expression. Many of the yeast intracellular signaling cascades have similar...
Ribosome Profiling
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...


