化学RAP通过RNA 5'基化解来发现特定的mRNA翻译调节
Hélène Ipas1, Ellen B Gouws1, Nathan S Abell1
1Department of Molecular Biosciences, University of Texas at Austin, 2500 Speedway, 78712, Austin, TX, USA.
EMBO reports
|January 23, 2024
概括
研究人员确定了在RNA上读取基化 (5'-Pme) 的蛋白质. 酶EPRS直接与5'-Pme结合,调节mRNA翻译和蛋白质定位,影响诸如利氏综合征等疾病.
科学领域:
- 分子生物学分子生物学
- 在RNA生物学,RNA生物学.
- 蛋白质组学是指蛋白质组学.
背景情况:
- RNA 5'-end 修改极大地影响RNA的稳定性和功能.
- 非正规的RNA封顶,如基甲基化 (5'-Pme),代表了一个新的调节机制.
- 了解与这些修饰帽子相互作用的蛋白质对于破译它们的生物作用至关重要.
研究的目的:
- 为了识别识别5'-PmeRNA修饰的细胞蛋白质.
- 阐明5'-Pme影响RNA-蛋白相互作用和基因表达的机制.
- 研究5'-Pme在特定疾病相关基因的作用.
主要方法:
- 化学RNA亲和性净化 (Chemical RNA Affinity Purification) 与定量蛋白质组学相结合,用于识别5'-Pme结合蛋白.
- 生物化学测试用于确认已识别的蛋白质和5'-PmeRNA之间的直接相互作用.
- 在具有改变5'-Pme编写活性的细胞中分析mRNA翻译和蛋白质定位.
主要成果:
- 核细胞链释放因子子1 (EPRS) 是多合成酶复合体 (MSC) 的核心组成部分,通过其链接域被确定为直接的5'-Pme读者.
- 5'-Pme编写器BCDIN3D被证明可以调节EPRS与特定mRNA的结合,特别是在启动密码子附近和富含MSC密码子的区域.
- BCDIN3D缺乏导致改变了EPRS结合,增加了LRPPRC的翻译和错位,LRPPRC是一种涉及法国加拿大利氏综合征的蛋白质.
结论:
- 这项研究揭示了EPRS作为5'-PmeRNA修饰的直接读者.
- BCDIN3D介导的RNA5'-Pme作为一个调控机制,控制EPRS-mRNA相互作用和随后的翻译.
- 这一途径突出了调节基因表达的新机制,这对理解与RNA修饰和蛋白质合成相关的疾病有意义.
更多相关视频
13:34Method for the Isolation and Identification of mRNAs, microRNAs and Protein Components of Ribonucleoprotein Complexes from Cell Extracts using RIP-Chip
Published on: September 29, 2012
27.6K
08:58In Vitro Transcribed RNA-based Luciferase Reporter Assay to Study Translation Regulation in Poxvirus-infected Cells
Published on: May 1, 2019
15.0K
相关概念视频
Regulation of Expression at Multiple Steps
910
The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
910
Regulation of Expression Occurs at Multiple Steps
22.7K
Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
22.7K
Regulated mRNA Transport
6.3K
In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing...
6.3K
RNA Stability
33.5K
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...
33.5K
Ribosome 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
Riboswitches
8.1K
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...
8.1K
