用修饰的寡核酸的化学合成及其酶性结合导致U6 SnRNA干环段的产生
Claudia Höbartner1, Ronald Micura
1Institute of Organic Chemistry, Leopold Franzens University, Innrain 52a, A-6020 Innsbruck, Austria.
Journal of the American Chemical Society
|January 30, 2004
概括
研究人员开发了标记的RNA用于X射线晶体学,使得精确的结构确定. 这种使用新型胺和酶结的方法对于理解复杂的RNA结构至关重要.
科学领域:
- 生物化学 生物化学
- 结构生物学 结构生物学
- 有机化学 有机化学
背景情况:
- 核酸的衍生物化对X射线晶体学具有前景.
- 多波长异常分散 (MAD) 技术从标签中受益.
- 之前的工作证明了这种方法在小DNA复合体上.
研究的目的:
- 开发和研究用于结构分析的标记RNA.
- 为了合成一种新型的2'-Se-methylcytidine phosphoramidite.
- 为了研究含有的RNA的结合和结合.
主要方法:
- 2'-Se-methylcytidine phosphoramidite的合成. 这是一个很好的方法.
- 聚氨酸核酸的固体相合成.
- 使用threo-1,4-dimercapto-2,3-butandiol (DTT) 的去保护.
- 使用T4RNA结合酶进行酶性结合.
主要成果:
- 成功合成并将新型胺酸纳入RNA.
- 有效地降低了标记RNA的保护和净化,长达22个月.
- 证明了含有的RNA的酶性结合,包括一个复杂的U6 snRNA干循环模式.
结论:
- 开发的方法可以为结构研究准备标记RNA.
- 标签方法是灵活的,并且与酶结相兼容.
- 这便于使用MAD.的复杂RNA分子的结构确定.
相关概念视频
Nucleotide Excision Repair
Overview
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...
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,...
RNA Editing
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...
Nucleotide Excision Repair
DNA Distortion and Damage
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...


