相关实验视频
Updated: Jul 6, 2026

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A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
Published on: September 15, 2021
一个活跃的I阶段spliceosome的分离及其RNP核心组成
Sergey Bessonov1, Maria Anokhina, Cindy L Will
1Department of Cellular Biochemistry, Max Planck Institute of Biophysical Chemistry, D-37077 Göttingen, Germany.
Nature
|March 7, 2008
概括
人类拼接酶可以在没有额外的蛋白质的情况下催化RNA拼接. 像Prp19-CDC5这样的关键蛋白质稳定了催化部位,揭示了结合体.
科学领域:
- 分子生物学分子生物学
- 在RNA生物学,RNA生物学.
- 生物化学 生物化学
背景情况:
- 蛋白质辅助对于在结合体中形成催化活性RNA结构至关重要.
- 结合体的活性部位所需的精确蛋白质组成在很大程度上是未知的.
研究的目的:
- 为了研究用于spliceosome催化活性的蛋白质要求.
- 为了识别建立和维持结合体的活性位点所必需的蛋白质.
主要方法:
- 人类结合体C复合体的亲和性净化.
- 在结合体激活过程中蛋白质组成变化的分析 (B到C复合体过渡).
- 破坏纯化的C复合体以隔离核心核核蛋白质 (RNP) 结构.
主要成果:
- 纯化的人类结合体C复合体独立表现出催化外链结合活性.
- 在B转C复合体过渡过程中发生了重要的蛋白质交换,稳定了Prp19-CDC5复合体蛋白质,破坏了SF3a/b蛋白质的稳定.
- 隔离了一个盐稳定的RNP核心,其中含有拼接中间体,U2,U5和U6小核RNA,以及U5和Prp19-CDC5蛋白质.
结论:
- 结合体具有内在的催化RNP域.
- Prp19-CDC5蛋白质和相关因素在维护结合体的催化活性结构方面发挥着核心作用.
相关概念视频
RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
Pre-mRNA Processing: RNA Splicing
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
Alternative RNA Splicing
Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
Ribosomal RNA Synthesis
Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...

