在转化DNA合成中,DNA聚合酶 κ 的催化和非催化功能
Selene Sellés-Baiget1, Sara M Ambjørn1, Alberto Carli2
1Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark.
Nature structural & molecular biology
|September 19, 2024
概括
聚合酶卡帕 (Polκ) 在转移DNA合成 (TLS) 中发挥双重作用,绕过DNA损伤. 它直接复制小沟病变,并非催化稳定大沟病变绕道复合体.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物化学 生物化学
背景情况:
- 转载DNA合成 (TLS) 对于复制具有病变的DNA至关重要.
- 聚合酶卡帕 (Polκ) 绕过小沟DNA附加物,但其在主要沟绕行中的作用尚不清楚.
- TLS是化疗的关键目标.
研究的目的:
- 阐明Polκ在绕过DNA损伤中的功能和调节.
- 为了研究Polκ在小沟和大沟DNA adduct bypass中的差异性作用.
- 了解涉及Rev1和PCNA无处不在的监管机制.
主要方法:
- 在人类细胞中使用CRISPR基编辑器选.
- 在Xenopus蛋提取物中转化DNA合成分析.
- 研究DNA病变和聚合酶相互作用.
主要成果:
- 波尔克在TLS中有两个不同的功能,由Rev1 binding规范.
- 波尔克对于绕过小沟病变至关重要,依赖PCNA无处不在,但独立于Rev1.
- 波尔克稳定了Rev1-Polζ复合体的主要沟损伤绕道,独立于其催化活性.
结论:
- 在转化DNA合成中,Polκ表现出催化和非催化功能.
- Rev1结合差异调节了Polκ在DNA损伤绕行中的作用.
- 这项研究揭示了Y家族TLS聚合酶的调节机制.
更多相关视频
11:08Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
Published on: June 19, 2018
9.7K
07:38DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
Published on: October 6, 2017
14.1K
相关概念视频
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
Proofreading
6.2K
Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore, it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase...
Errors During Replication are Corrected by the DNA Polymerase...
6.2K
DNA Topoisomerases
31.1K
Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
Types and Mechanism of action
Topoisomerases are divided into two main types. ...
Types and Mechanism of action
Topoisomerases are divided into two main types. ...
31.1K
The Replisome
33.2K
DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
33.2K
Replication in Eukaryotes
170.4K
Overview
170.4K
Homologous Recombination
50.3K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
50.3K
