使用体外选择阐明子头利酶的进化机制和变体
Jake Brill1, Connor Nurmi1, Yingfu Li1
1Department of Biochemistry and Biomedical Sciences, McMaster University, 1280 Main Street West, Hamilton, Ontario, L8S 4K1, Canada.
Chembiochem : a European journal of chemical biology
|August 8, 2024
概括
通过系统突变和体外选择发现新的Hammerhead Ribozyme (HHR) 变体,揭示了必需的核酸可以耐受变化,扩大了我们对RNA酶进化和功能的理解.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 在RNA催化过程中,RNA催化
背景情况:
- 头 ribozyme (HHR) 是一种关键的RNA酶,催化特定的分裂反应.
- 传统上,人们认为HHR催化核中的突变会消除活性.
- 最近对自然变异的发现挑战了这一观点,表明了复杂的进化格局.
研究的目的:
- 为了探索全长头Ribozyme的序列要求和进化途径.
- 研究突变对HHR催化核活性的影响.
- 识别新的HHR变体并了解它们的功能意义.
主要方法:
- 在HHR催化核心中系统地引入单,双和三重突变.
- 创建一个全面的突变库.
- 在体外选择以确定活跃的 ribozyme 变体.
主要成果:
- 识别了许多新的Hammerhead Ribozyme变体.
- 在以前必不可少的核酸中发现具有突变的变体.
- 证明核酸进化轨迹与功能重要性之间的相关性.
结论:
- 头Ribozyme的催化核心表现出比以前假设的更大的突变耐受性.
- 核酸的进化史可以作为核酸功能重要性的一个预测指标.
- 这些发现为HHR演变提供了新的见解,并提供了评估功能核酸序列要求的潜在方法.
相关概念视频
Ribozymes
11.2K
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...
Ribozymes can...
11.2K
Conservative Site-specific Recombination and Phase Variation
6.0K
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...
6.0K
Catalytically Perfect Enzymes
3.9K
The theory of catalytically perfect enzymes was first proposed by W.J. Albery and J. R. Knowles in 1976. These enzymes catalyze biochemical reactions at high-speed. Their catalytic efficiency values range from 108-109 M-1s-1. These enzymes are also called 'diffusion-controlled' as the only rate-limiting step in the catalysis is that of the substrate diffusion into the active site. Examples include triose phosphate isomerase, fumarase, and superoxide dismutase.
Most enzymes...
Most enzymes...
3.9K
Bacterial RNA Polymerase
29.4K
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...
29.4K
Gene Evolution - Fast or Slow?
7.1K
The genomes of eukaryotes are punctuated by long stretches of sequence which do not code for proteins or RNAs. Although some of these regions do contain crucial regulatory sequences, the vast majority of this DNA serves no known function. Typically, these regions of the genome are the ones in which the fastest change, in evolutionary terms, is observed, because there is typically little to no selection pressure acting on these regions to preserve their sequences.
In contrast, regions which code...
In contrast, regions which code...
7.1K
In-vitro Mutagenesis
13.9K
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
13.9K


