NTPs在RNA聚合酶I和II的活性位点上进行竞争
Kaila B Fuller1, Ryan M Requijo1, David A Schneider2
1Department of Chemistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Biophysical chemistry
|August 24, 2024
概括
研究人员开发了一种新的分析方法来研究RNA聚合酶 (Pols). 这种方法揭示了非相关核酸与相关核酸竞争,为转录能量和选择性提供了洞察力.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 类真核生物利用三种RNA聚合酶 (Pols) 来进行转录,而细菌和古生物则只使用一种.
- 了解这些极点对核酸添加的动力机制对于破译转录忠实性至关重要.
研究的目的:
- 开发和验证RNA聚合酶动力学中意外延伸产物的分析策略.
- 在真核生物Pols.的活性部位中研究同源和非同源核酸之间的热力学竞争.
主要方法:
- 进行过渡状态动力学实验以分析单核和多核酸添加.
- 开发了一种新的分析策略,以考虑意想不到的扩展产品,包括错误整合和NTP污染.
主要成果:
- 开发的分析策略成功地考虑了意想不到的扩展产品.
- 发现了证据,表明非相关核酸和相关核酸之间的热力学竞争对Pol I,ΔA12 Pol I和Pol II的活性部位.
- 这种竞争性结合表明,相关的和非相关的核酸亲和关系处于一个数量级之内.
结论:
- 新的分析方法可以提取关于核酸进入能量和选择性的额外信息.
- 这些发现挑战了仅基于基配对能量学的绝对选择性假设.
- 这项工作为控制转录精度的复杂机制提供了更深入的见解.
相关概念视频
Transcription Initiation
16.3K
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...
16.3K
RNA Polymerase II Accessory Proteins
9.1K
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...
9.1K
Eukaryotic RNA Polymerases
24.0K
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...
24.0K
Bacterial RNA Polymerase
29.4K
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
29.4K
Translesion DNA Polymerases
9.9K
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
9.9K
Bacterial Transcription
28.1K
RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:
Transcription can be divided into three main stages, each involving distinct DNA sequences to guide the polymerase. These are:
28.1K


