使用突变概况的tRNA修饰酶的功能分析
Ryota Yamagami1, Hiroyuki Hori1
1Department of Materials Science and Biotechnology, Graduate School of Science and Engineering, Ehime University, Matsuyama, Ehime, Japan.
Methods in enzymology
|November 4, 2023
概括
这项研究提出了一种高通量突变分析方法,用于分析tRNA甲基转移酶酶的酶功能. 该协议使用TrmK进行了演示,为各种tRNA修饰提供了对基质识别的机制性见解.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 遗传学 是一个遗传学.
背景情况:
- 转移RNA (tRNA) 对于蛋白质合成至关重要,作为传递 RNA (mRNA) 编码码解码的适配分子.
- 转录后的修改对于tRNA结构,功能以及与酶和其他分子的相互作用至关重要.
- 特定站点的tRNA修饰酶催化了大多数修饰核酸的合成,影响了各种生物过程.
研究的目的:
- 提供使用突变分析的tRNA甲基转移酶酶功能高通量分析的详细协议.
- 通过使用TrmK作为模型酶,获得tRNA甲基转移酶对基质识别的机制性见解.
主要方法:
- 开发和应用一个突变分析协议.
- 使用Geobacillus stearothermophilus的tRNA m1A22甲基转移酶TrmK作为一个模型系统.
- 酶基质相互作用的高通量分析.
主要成果:
- 证明了突变分析对研究tRNA甲基转移酶机制的有用性.
- 提供了有关TrmK如何识别其基质tRNA的机制性见解.
- 建立了适用于各种tRNA修饰酶的多功能协议.
结论:
- 开发的突变分析协议能够对tRNA甲基转移酶活性进行高通量分析.
- 这种方法为tRNA修饰酶及其基质识别提供了有价值的机械洞察力.
- 该协议可适应研究涉及对tRNA的沃森-克里克面部修改的一系列酶.
关键词:
高通量选的高通量选突变概况分析 (mutational profiling) 是一种关于突变概况的分析.的RNA甲基转移酶.基因组RNA的修饰是RNA的修饰.RNA与蛋白质的相互作用转移RNA转移RNA是什么意思更多相关视频
09:04Studying Ribonucleotide Incorporation: Strand-specific Detection of Ribonucleotides in the Yeast Genome and Measuring Ribonucleotide-induced Mutagenesis
Published on: July 26, 2018
7.8K
11:36A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
Published on: July 3, 2016
10.9K
相关概念视频
02:36Transfer RNA Synthesis
12.0K
One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
12.0K
02:26tRNA Activation
19.3K
Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
19.3K
01:36Mismatch Repair
40.2K
Overview
40.2K
02:24Ribosome Profiling
3.5K
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...
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...
3.5K
01:18Mutations in Microorganisms
18
Mutations are heritable changes in an organism’s genome involving alterations in the base sequence of DNA or RNA. These changes can influence cellular processes and phenotypic traits, potentially transforming the unaltered wild type into a mutant form. Such changes, termed forward mutations, are pivotal in shaping the genetic diversity of organisms.RNA viruses exhibit the highest mutation rates due to the absence of robust proofreading mechanisms during genome replication. In contrast,...
18