相关实验视频
Updated: Jun 3, 2026

09:12
DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
Published on: September 16, 2019
RNA聚合酶II的C端域通过特定位点的甲基化来改变
Robert J Sims1, Luis Alejandro Rojas1, David B Beck1
1Howard Hughes Medical Institute (HHMI), Department of Biochemistry, New York University School of Medicine, 522 First Avenue, Smilow 211, New York, NY 10016, USA.
概括
RNA聚合酶II (RNAPII) 的CTD甲基化由CARM1在R1810之前的转录启动. 这种甲基化对于特定小RNAs的表达至关重要,影响基因调节.
科学领域:
- 分子生物学分子生物学
- 基因表达 基因表达
- 翻译后修改 翻译后修改
背景情况:
- RNA聚合酶II (RNAPII) 的碳氧终端域 (CTD) 在翻译后被广泛修改.
- 这些修改对于调节转录启动和延长至关重要.
研究的目的:
- 调查阿尔金因甲基化在RNAPIICTD修饰中的作用.
- 确定负责CTD甲基化的特定酶及其功能后果.
主要方法:
- 在体外酶分析以确定CARM1在RNAPIICTD上的活性.
- 在RNAPII中对R1810的局部定向突变发生.
- 在野生类型和突变细胞/纤维细胞中分析小RNA表达.
主要成果:
- 同活性剂相关的阿尔金因甲基转移酶1 (CARM1) 在阿尔金因1810 (R1810) 中甲基化RNAPII CTD.
- 甲基化在活体中发生在高酸化RNAPII上,但在体外被Ser2/Ser5酸化抑制,这表明了启动前的时间.
- R1810突变或CARM1缺乏导致小核RNA和小核RNA的错误表达.
结论:
- 在R1810处由CARM1进行CTD甲基化是一种RNAPII的新型后翻译修饰.
- 这种甲基化事件对于选择小RNA的正确表达至关重要.
- CTD甲基化可能在为不同基因类型招募特定的转录机制中发挥作用.
相关概念视频
Eukaryotic RNA Polymerases
RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
All three eukaryotic RNAPs require specific transcription factors, of which the...
Eukaryotic RNA Polymerases
RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
All three eukaryotic RNAPs require specific transcription factors, of which the...
Transcription Initiation
Initiation is the first step of transcription in eukaryotes. Prokaryotic RNA Polymerase (RNAP) can bind to the template DNA and start transcribing. On the other hand, transcription in eukaryotes requires additional proteins, called transcription factors, to first bind to the promoter region in the DNA template. This binding helps recruit the specific RNAP that can assemble on the DNA and start transcription.
The promoters and enhancers and their accessory proteins allow tight regulation of...
The promoters and enhancers and their accessory proteins allow tight regulation of...
RNA Polymerase II Accessory Proteins
Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
RNA Polymerase II Accessory Proteins
Proteins that regulate transcription can do so either via direct contact with RNA Polymerase or through indirect interactions facilitated by adaptors, mediators, histone-modifying proteins, and nucleosome remodelers. Direct interactions to activate transcription is seen in bacteria as well as in some eukaryotic genes. In these cases, upstream activation sequences are adjacent to the promoters, and the activator proteins interact directly with the transcriptional machinery. For example, in...
Transfer RNA Synthesis
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...

