特定位点 ((II) 交叉链接在一个 ribozyme 活性位点
Erich G Chapman1, Victoria J DeRose
1Department of Chemistry and Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403, USA.
Journal of the American Chemical Society
|November 11, 2011
概括
研究人员开发了一种新的(II) RNA-RNA交叉链接方法,使用酸替代物. 这种技术使得RNA的特定位点结构分析成为可能,这在Hammerhead ribozyme中得到了证明.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 结构生物学 结构生物学
背景情况:
- RNA结构和动态对于功能至关重要.
- 特定站点的交叉链接为RNA结构生物学提供了有价值的距离约束.
- 现有的方法需要进一步改进,以便精确地确定RNA的结构.
研究的目的:
- 引入一种新的RNA-RNA交联策略,利用 (II) 复合体.
- 为了研究这种方法在探测Hammerhead ribozyme的结构中的应用.
- 建立一个新的工具,用于特定地点的RNA结构和动态研究.
主要方法:
- 一个新的RNA-RNA交叉链路策略,基于Pt(II) 向酸替代物.
- 动态招募和定cis-diammine Pt(II) 复合物到酸替代RNA.
- 通过Pt(II) 复合置换诱导RNA核基之间的金属介导交联.
主要成果:
- Pt(II) 交叉链接发生在Hammerhead ribozyme活性部位内的G8和G10核酸中,当酸在分裂键处时.
- 交联效率高达30%的产量取决于Mg2+) 离子度和RNA三级结构的形成.
- 交叉链接被观察到在活性位点内,而不是在RNA螺旋中,这表明该位点的特异性.
结论:
- 开发的基于 Pt (II) 的交叉链路策略对于特定站点的RNA结构分析是有效的.
- 这种方法为探索RNA结构和动态提供了一个有前途的新工具,特别是在复杂的RNA基因中.
- 这些发现突出了动态惰性Pt(II) 复合体在促进RNA结构生物学方面的潜力.
相关概念视频
Ribozymes
The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
Ribozymes can be...
Ribozymes
The term ribozyme is used for RNA that can act as an enzyme. Ribozymes are mainly found in selected viruses, bacteria, plant organelles, and lower eukaryotes. Ribozymes were first discovered in 1982 when Tom Cech’s laboratory observed Group I introns acting as enzymes. This was shortly followed by the discovery of another ribozyme, Ribonulcease P, by Sid Altman’s laboratory. Both Cech and Altman received the Nobel Prize in chemistry in 1989 for their work on ribozymes.
Ribozymes can be...
Ribozymes can be...
Riboswitches
Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
Ligand Binding and Linkage
Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked. In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence the...
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...
Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...


