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相关概念视频

Proofreading01:31

Proofreading

6.3K
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...
6.3K
Mismatch Repair01:20

Mismatch Repair

4.9K
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
4.9K
Overview of DNA Repair02:25

Overview of DNA Repair

31.0K
In order to be passed through generations, genomic DNA must be undamaged and error-free. However, every day, DNA in a cell undergoes several thousand to a million damaging events by natural causes and external factors. Ionizing radiation such as UV rays, free radicals produced during cellular respiration, and hydrolytic damage from metabolic reactions can alter the structure of DNA. Damages caused include single-base alteration, base dimerization, chain breaks, and cross-linkage.
Chemically...
31.0K
Base Excision Repair01:54

Base Excision Repair

22.4K
One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
The first step of...
22.4K
Genome Copying Errors02:46

Genome Copying Errors

4.2K
DNA replication is a well-evolved process that copies millions of base pairs with high fidelity during each cell division. Occasionally a wrong base or a long stretch of wrong bases may get added to the daughter strands. If the errors are left unchecked, cells might accumulate several mutations that might endanger their  survival. Therefore, the copying errors are checked and repaired at three levels.
4.2K
Base-pairing and DNA Repair02:27

Base-pairing and DNA Repair

64.8K
64.8K

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相关实验视频

Updated: Jul 7, 2025

Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis

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对于DNA校对的结构基础

Gina Buchel1, Ashok R Nayak1, Karl Herbine1

  • 1Department of Biochemistry and Molecular Biology, Thomas Jefferson University, 1020 Locust St, Philadelphia, PA, 19107, USA.

Nature communications
|December 27, 2023
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概括

人类线粒体DNA聚合酶 (Polγ) 的校对涉及一个保存的"螺栓作用"机制. 这个过程反复地将聚合酶沿着DNA转移,而不会发生解离,从而使错误纠正对细胞活力至关重要.

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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis

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Atomic Force Microscopy Investigations of DNA Lesion Recognition in Nucleotide Excision Repair
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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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科学领域:

  • 分子生物学分子生物学
  • 生物化学 生物化学
  • 遗传学 是一个遗传学.

背景情况:

  • DNA聚合酶 (DNAP) 校对对于保持基因组完整性和细胞活力至关重要.
  • 在DNAP校对过程中错误转移和DNA重新定位的精确机制仍然不完全理解.

研究的目的:

  • 阐明由人类线粒体DNA聚合酶 (Polγ) 进行DNA校对的逐步机制.
  • 揭示涉及从聚合酶到外核酶部位转移不匹配的结构动态.

主要方法:

  • 在校对过程中,高分辨率结构分析了人类Polg的九个不同的状态.
  • 功能测定和突变发生研究,以验证拟议的机制.

主要成果:

  • 详细的结构捕捉了关键事件:不匹配的基因识别,与聚合酶部位的解离,DNAP转位,DNA轨迹的变化和原料重新定位.
  • 有证据表明"螺栓作用"机制涉及代DNAP转位而无解离,从而促进了原料转移.
  • 病原性突变与涉及校对步骤的关键结构元素有关.

结论:

  • 一个保存的"螺栓作用"机制控制了人类Polγ的DNA校对,涉及连续的聚合酶转位.
  • 这种机制确保了有效的错误纠正,并突出了特定结构元素的功能重要性.
  • 了解这一过程,可以了解DNA复制的真实性和相关的遗传疾病.