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関連する概念動画

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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関連する実験動画

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

DNAポリメラーゼが正しいものと間違ったものを選択することを観察する.

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
まとめ
この要約は機械生成です。

DNAポリメラーゼβは,リアルタイムで触媒中間物質を明らかにするために,天然の基板を使用します. 新しい構造は,活性部位の調整がDNA合成の精度とゲノムの安定性を改善する方法を示しています.

さらに関連する動画

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
07:38

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

Published on: October 6, 2017

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

関連する実験動画

Last 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

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
07:38

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

Published on: October 6, 2017

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

科学分野:

  • バイオケミストリー バイオケミストリー
  • 構造生物学 構造生物学とは
  • 分子生物学は分子生物学である.

背景:

  • DNAポリメラーゼβ (polβ) は,DNA修復と隙間補充合成に不可欠である.
  • カタリシス中のニュクレオチジル転送には2つの金属イオンを使用します.
  • 以前の研究では,触媒中間物質を捕獲するために基板類型に依存していました.

研究 の 目的:

  • DNAポリメラーゼβ活動中の新しい触媒中間物質を特定する.
  • ポリメラーゼの忠誠性とゲノム安定性の基礎となる分子機構を理解する.
  • 天然基板を用いてDNA合成における金属イオンの役割を調査する.

主な方法:

  • DNAポリメラーゼβ反応のために使用された天然基板 (正しいおよび不正な核酸).
  • 製品形成をリアルタイムで捉えるために,15の異なる結晶構造を決定しました.
  • ヌクレオチド挿入中の活性部位の構造変化を分析した.

主要な成果:

  • 正確なヌクレオチド挿入を好み,誤った挿入を好まないダイナミックな活性部位の調整が観察されました.
  • 正確なヌクレオチド組み込み時にのみ形成される一時的な第3金属結合部位を特定しました.
  • ピロホスファートの解離は,サブドメインの再定位に関連した,誤ったヌクレオチド挿入の後,より速いことが判明しました.

結論:

  • この研究は,DNAポリメラーゼβ触媒における,これまで認識されていなかった分子調整を明らかにした.
  • 暫定的な第3の金属部位とサブドメインの動きは,ポリメラーゼの忠誠性に寄与する.
  • これらの発見は,正確なDNA合成を通じてゲノムの安定性を維持するための洞察を提供します.