相关实验视频
Updated: Nov 18, 2025

07:02
An Assay for Quantifying Protein-RNA Binding in Bacteria
Published on: June 12, 2019
6.8K
细胞中的RNA结合蛋白的动态景观
Deepak Sharma1,2, Leah L Zagore1,2, Matthew M Brister3
1Center for RNA Science and Therapeutics, School of Medicine, Case Western Reserve University, Cleveland, OH, USA.
Nature
|February 11, 2021
概括
研究人员开发了一种新方法来测量RNA结合蛋白 (RBPs) 附着和脱离细胞中的RNA的速度. 这种KIN-CLIP技术揭示了快速结合动态,将RBP相互作用与基因表达调节联系起来.
科学领域:
- 分子生物学
- 遗传学
- 生物化学
背景情况:
- 基因表达涉及RNA结合蛋白 (RBPs) 和真核细胞中的RNA分子之间的复杂相互作用.
- 了解RBP-RNA结合的动力学对于协调细胞RNA处理和功能至关重要.
- 在活细胞中测量这些动力参数是一个重大的挑战.
研究的目的:
- 开发和验证一种用于实时测量细胞内单个RNA位点的RBP结合和解离动力学的新方法.
- 通过这种新方法研究RBP DAZL的结合动态.
- 将RBP结合动力学与其对mRNA水平和翻译的功能影响相关联.
主要方法:
- 开发动力交叉链接和免疫沉 (KIN-CLIP),使用时间解析的RNA-蛋白交叉链接与脉冲的秒紫外线激光.
- 应用免疫沉和高通量测序来量化RBP结合率和解离率.
- 对RBP DAZL数千个单独的RNA结合位点进行分析.
主要成果:
- 通过KIN-CLIP方法,成功地确定了RBP DAZL在细胞中数千个RNA位点的结合和解离动力学.
- 发现DAZL与单个RNA位点的结合时间非常短 (数秒或更短),解离时间显著更长.
- DAZL结合通常发生在近位中,整体mRNA调节与这些内的累积结合概率相关.
结论:
- 现在可以在活细胞中实验测量RNA-蛋白相互作用的动态参数.
- 该研究提供了RBP-RNA结合动力学,聚类行为和基因表达的调节之间的定量联系.
- 了解RBP结合动态对于解读它们在细胞RNA代谢和功能中的作用至关重要.
相关概念视频
Types of RNA
70.8K
Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
70.8K
Nucleic Acids
47.9K
Nucleic acids are the most important macromolecules for the continuity of life. They carry the cell's genetic blueprint and carry instructions for its functioning.
DNA and RNA
The two main types of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). DNA is the genetic material in all living organisms, ranging from single-celled bacteria to multicellular mammals. It is in the nucleus of eukaryotes and in the organelles, chloroplasts, and mitochondria. In prokaryotes,...
DNA and RNA
The two main types of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). DNA is the genetic material in all living organisms, ranging from single-celled bacteria to multicellular mammals. It is in the nucleus of eukaryotes and in the organelles, chloroplasts, and mitochondria. In prokaryotes,...
47.9K
Nucleic acids
184.5K
Nucleic acids are the most important macromolecules for the continuity of life. They carry the cell's genetic blueprint and carry instructions for its functioning.
DNA and RNA
The two main types of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). DNA is the genetic material in all living organisms, ranging from single-celled bacteria to multicellular mammals. It is in the nucleus of eukaryotes and in the organelles, chloroplasts, and mitochondria. In prokaryotes,...
DNA and RNA
The two main types of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). DNA is the genetic material in all living organisms, ranging from single-celled bacteria to multicellular mammals. It is in the nucleus of eukaryotes and in the organelles, chloroplasts, and mitochondria. In prokaryotes,...
184.5K
Bacterial RNA Polymerase
31.6K
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...
31.6K
RNA Stability
34.8K
Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
34.8K
Cooperative Binding of Transcription Regulators
6.9K
Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome. Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form...
6.9K

