相关实验视频

Updated: Jun 30, 2025

Atomic Force Microscopy Investigations of DNA Lesion Recognition in Nucleotide Excision Repair

Atomic Force Microscopy Investigations of DNA Lesion Recognition in Nucleotide Excision Repair

Published on: May 24, 2017

对AP内核酶AtARP的催化机制的结构洞察力

Wenting Guo¹, Weijun Wu¹, Yan Wen²

¹Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Guangdong-Hong Kong Joint Laboratory for RNA Medicine, Medical Research Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, China.

Structure (London, England : 1993)

|March 19, 2024

概括

这项研究揭示了Arabidopsis AP内核酶AtARP的晶体结构,详细介绍了它在植物DNA修复和基切除修复 (BER) 中的作用. 这些发现提供了关于植物DNA脱甲基和基因组防御机制的见解.

科学领域:

分子生物学分子生物学
遗传学遗传学是一个
生物化学生物化学

背景情况:

基切除修复 (BER) 是修复来自不同来源的DNA损伤的关键途径.
AP内核酶是BER中的关键酶,负责处理基底部位.
在植物中,BER参与了5-甲基细胞素 (5mC) 的活性DNA脱甲基化.

研究的目的:

为了确定Arabidopsis AP内核酶 (AtARP) 与DNA结合的晶体结构,模仿一个底层位点.
通过AtARP阐明基底部部位识别和裂变的分子机制.
将AtARP与其他植物AP内核酶进行比较,并评估其在异质系统中的DNA修复功能.

主要方法:

进行X射线晶体学,以获得AtARP-DNA复合体的高分辨率结构.
位点定向突变发生,以确定参与DNA结合和催化过程的关键氨基酸残留物.
在哺乳动物细胞系中进行功能性测试,以评估AtARP的DNA修复能力.

主要成果:

晶体结构揭示了AtARP如何在含有四基 (THF) 的基底部模拟点上结合和分裂DNA.
特定的残留物被确定为基底部结合,去除和孤儿基相互作用至关重要.
对比分析突出了植物AP内核酶的独特特征,AtARP证明了哺乳动物细胞中的DNA修复活性.

结论:

这项研究为植物基切除修复提供了详细的机制性见解,特别是AtARP的作用.
这些发现有助于理解植物的DNA修复和表观遗传调节.
AtARP的保存功能表明在跨物种基因组稳定性研究中的潜在应用.

关键词:

5 - 甲基氨酸 (Methycytosine) 的使用在AP内核酶中使用.在ARP中,ARP是ARP.阿拉比多普西斯 (Arabidopsis) 是一种植物.造成的DNA损伤是DNA损伤.SAP 域名 SAP 域名基础切除修复的基础切除修复

更多相关视频

Single-Molecule Dwell-Time Analysis of Restriction Endonuclease-Mediated DNA Cleavage

Single-Molecule Dwell-Time Analysis of Restriction Endonuclease-Mediated DNA Cleavage

Published on: February 7, 2021

Application of Stopped-flow Kinetics Methods to Investigate the Mechanism of Action of a DNA Repair Protein

Application of Stopped-flow Kinetics Methods to Investigate the Mechanism of Action of a DNA Repair Protein

Published on: March 31, 2010

相关概念视频

Allosteric Proteins-ATCase

Allosteric Proteins-ATCase

Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...

Restarting Stalled Replication Forks

Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...

Long-patch Base Excision Repair

Long-patch Base Excision Repair

Since the discovery of the two BER pathways, there has been a debate about how a cell chooses one pathway over the other and the factors determining this selection. Numerous in vitro experiments have pointed out multiple determinants for the sub-pathway selection. These are:

Generation of Straight or Branched Actin Filaments

Generation of Straight or Branched Actin Filaments

The straight or branched structure formation of actin filaments is controlled by nucleating proteins such as the formins and Arp2/3 complex. Formin-mediated assembly results in straight filaments, whereas Arp2/3 protein complex-mediated assembly results in branched actin filaments.
Arp2/3 Complex
Arp2/3 complex is a seven-subunit complex consisting of two proteins similar to actin- Arp2 and Arp3, and five other subunits that help keep Arp2 and Arp3 inactive. When required, the complex is...

Eukaryotic RNA Polymerases

Eukaryotic RNA Polymerases

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...

ATP Synthase: Mechanism

ATP Synthase: Mechanism

In animals, the mitochondrial F1F0 ATP synthase is the key protein that synthesizes ATP molecules through a complex catalytic mechanism. While the nuclear genome encodes the majority of ATP synthase subunits, the mitochondrial genome encodes some of the enzyme's most critical components. The formation of this multi-subunit enzyme is a complex multi-step process regulated at the level of transcription, translation, and assembly. Defects in one or more of these steps can result in decreased...