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相关概念视频

Proofreading01:31

Proofreading

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 Enzyme
Proofreading01:43

Proofreading

Overview
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

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...
DNA Replication02:40

DNA Replication

DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied.  After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
Replication in Prokaryotes
DNA replication uses a large number of...
Homologous Recombination02:31

Homologous Recombination

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...
Replication in Eukaryotes02:31

Replication in Eukaryotes

Overview

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相关实验视频

Updated: May 10, 2026

Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
11:08

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

Published on: June 19, 2018

观察DNA聚合酶从错误中选择正确的

Bret D Freudenthal1, William A Beard, David D Shock

  • 1Laboratory of Structural Biology, National Institute of Environmental Health Sciences, National Institutes of Health, P.O. Box 12233, Research Triangle Park, NC 27709-2233, USA.

Cell
|July 6, 2013
PubMed
概括
此摘要是机器生成的。

DNA聚合酶β使用自然基质来揭示实时的催化中间体. 新的结构显示了活点调整如何提高DNA合成精度和基因组稳定性.

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Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay
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Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay

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Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
11:08

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

Published on: June 19, 2018

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
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DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

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Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay
17:03

Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay

Published on: March 23, 2010

科学领域:

  • 生物化学 生物化学
  • 结构生物学 结构生物学
  • 分子生物学分子生物学

背景情况:

  • DNA聚合酶β (pol β) 对于DNA修复和填补缺口的合成至关重要.
  • 它在催化过程中利用两个金属离子进行核基转移.
  • 之前的研究依赖基质类似物来捕获催化中间体.

研究的目的:

  • 在DNA聚合酶β活性过程中识别新的催化中间体.
  • 了解构成聚合酶忠实性和基因组稳定性的分子机制.
  • 用天然基质研究金属离子在DNA合成中的作用.

主要方法:

  • 用于DNA聚合酶β反应的自然基质 (正确和不正确的核酸).
  • 确定了15个不同的晶体结构以捕捉实时产品形成.
  • 在核酸插入过程中分析了活性部位的结构变化.

主要成果:

  • 观察到动态活性部位调整有利于正确的核酸插入,不利于不正确的插入.
  • 确定了一个短暂的第三金属结合点,仅在正确的核酸结合过程中形成.
  • 发现核酸盐解离在错误的核酸插入后更快,这与子域重新定位有关.

结论:

  • 这项研究揭示了DNA聚合酶β催化过程中以前未被认可的分子调整.
  • 一个短暂的第三金属位点和子域运动有助于聚合酶的忠实性.
  • 这些发现为通过精确的DNA合成来维持基因组稳定提供了洞察力.