单子单元RNA聚合酶,KpnP,Ro45Iw和CD23823,具有精确的终端合成
Haruka Takatsuki1, Ryota Miyachi2, Kaito Seo3
1College of Arts and Science, The University of Tokyo, Meguro, Tokyo, Japan.
The Journal of biological chemistry
|June 25, 2025
概括
包括CD23823在内的新菌体RNA聚合酶 (RNAPs) 提供了比T7RNAP更好的RNA合成精度. 这些酶为苛刻的应用提供了更均的5'和3'终端.
科学领域:
- 分子生物学分子生物学
- 酶学 是一种酶学.
- 生物技术是生物技术.
背景情况:
- T7RNA聚合酶 (RNAP) 是用于体外RNA合成的广泛使用的酶,但产生具有异质3'-末端的RNA.
- 现有的替代RNAP存在局限性,不能完全取代T7RNAP.
- 需要具有精确RNA合成能力的新型RNAP.
研究的目的:
- 为了识别和表征新的菌体RNA聚合酶 (RNAPs),以改善体外RNA合成.
- 为了评估RNA合成活动,最佳温度和新RNAPs的盐耐受性.
- 与T7RNAP相比,评估这些新型RNAP产生的5'和3'终端的精度.
主要方法:
- 对菌体RNAP数据库的选,以识别候选聚合酶及其促进体.
- 选择的RNAP的重组表达和特征 (KpnP,Ro45Iw,CD23823). 它们的重组表达和特征.
- 通过深度测序评估RNA合成活性,最佳温度,盐耐受性和5'/3'末端均性.
主要成果:
- 三种新型RNAP (KpnP,Ro45Iw,CD23823) 被确定并进行了表征.
- 这些RNAP的RNA合成活性与T7RNAP的RNA合成活性相似,偏好较低的温度.
- CD23823表现出更高的盐分耐受性,并且产生了比T7 RNAP更同质的RNA终端.
结论:
- 之前未被描述的菌体RNAP,特别是CD23823,是T7RNAP的合适替代品.
- 这些新型RNAP在RNA5'和3'末端合成中提供了更高的精度.
- CD23823是RNA合成应用的有前途的酶,需要明确的术语.
相关概念视频
Bacterial RNA Polymerase
30.6K
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...
30.6K
Eukaryotic RNA Polymerases
24.8K
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...
24.8K
Transcription Initiation
16.9K
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...
16.9K
Translesion DNA Polymerases
10.2K
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...
10.2K
Lagging Strand Synthesis
54.4K
During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
54.4K
Bacterial Transcription
29.8K
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:
29.8K


