重新设计tRNA甲基转移酶以共价捕获新的RNA基质
Tyler S Smith1,2, Madeline A Zoltek1,2, Matthew D Simon1,2
1Department of Molecular Biophysics & Biochemistry , Yale University , New Haven , Connecticut 06511 , United States.
Journal of the American Chemical Society
|October 19, 2019
概括
研究人员设计了RNA修饰酶,以创建研究RNA的新工具. 他们开发了一种突变酶,可以被特定的RNA基质共聚地捕获,从而开辟新的研究途径.
科学领域:
- 生物化学
- 分子生物学
- 核糖核酸生物学
背景情况:
- 对RNA功能和新陈代谢至关重要.
- 安装RNA修饰的酶是开发研究工具的关键目标.
- 了解这些酶的基质特异性对于它们的设计至关重要.
研究的目的:
- 探索尿素-54tRNA甲基转移酶 (TrmA) 的可改造性,以改变其基质特异性.
- 通过酶工程开发研究和操纵RNA的新工具.
- 探测TrmA基质的可塑性.
主要方法:
- 使用TrmA突变体 (TrmA*),该突变体与基质RNA保持共价结合.
- 采用合理设计和高通量测序试验来分析TrmA*RNA基质.
- 使用高通量基质选选额外的TrmA*突变物.
主要成果:
- 合理工程的TrmA*显著改变了它的基质特异性.
- 工程基质突变物与野生类型的TrmA保持活动.
- 鉴定了一种三重突变RNA基质,该基质被一种工程化TrmA*双重突变分子有效地捕获,但不是野生型TrmA.
结论:
- 蛋白质工程可以重新配置RNA修饰酶的基质特异性.
- 这项研究为开发可共捕获特定RNA的工程蛋白质提供了基础.
- 开发的工具为RNA研究和操纵提供了新的可能性.
相关概念视频
Transfer RNA Synthesis
13.1K
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...
13.1K
Transfer RNA Synthesis
3.5K
3.5K
RNA Editing
9.7K
RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
9.7K
tRNA Activation
22.3K
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...
22.3K
tRNA Activation
8.2K
8.2K
Types of RNA
8.8K
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 regulating 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 Performs Diverse...
RNA Performs Diverse...
8.8K


