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相关概念视频

Mismatch Repair01:36

Mismatch Repair

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...
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...
Restarting Stalled Replication Forks02:37

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, a...
Mismatch Repair01:20

Mismatch Repair

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...
Phase II Reactions: Methylation Reactions01:17

Phase II Reactions: Methylation Reactions

Methylation is a phase II biotransformation process involving the attachment of a methyl group to a substrate. Enzymes known as methyltransferases orchestrate this reaction.
The mechanism of methylation unfolds in two stages. The first stage sees a methyltransferase enzyme facilitating the transfer of a methyl group from S-adenosylmethionine (SAM) to the substrate, forming S-adenosylhomocysteine (SAH). The second stage involves further metabolism of SAH into homocysteine, which can be recycled...

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相关实验视频

Updated: May 12, 2026

DamID-seq: Genome-wide Mapping of Protein-DNA Interactions by High Throughput Sequencing of Adenine-methylated DNA Fragments
09:14

DamID-seq: Genome-wide Mapping of Protein-DNA Interactions by High Throughput Sequencing of Adenine-methylated DNA Fragments

Published on: January 28, 2016

从非特异性的DNA相互作用过渡到特定的DNA相互作用沿着大甲基转移酶的基质识别途径.

John R Horton1, Kirsten Liebert, Stanley Hattman

  • 1Department of Biochemistry, Emory University School of Medicine, 1510 Clifton Road, Atlanta, GA 30322, USA.

Cell
|May 11, 2005
PubMed
概括

对DNA甲基转移酶的结构研究揭示了这些酶如何实现DNA序列的特异性. 研究人员确定了对有效和选择性DNA甲基化至关重要的歧视性和反歧视性接触.

科学领域:

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

背景情况:

  • DNA 甲基转移酶 (DMTases) 是催化甲基转移到 DNA 基的酶.
  • 这些酶识别并与特定的DNA序列结合以进行甲基化,这是一个关键的表观遗传修饰.
  • 了解DMTases对DNA序列识别的分子机制对于破译基因调节至关重要.

研究的目的:

  • 阐明通过DNA甲基转移酶识别DNA序列的结构基础.
  • 研究特定氨基酸残留在区分目标DNA序列中的作用.
  • 了解甲基化过程中从非特异性转变为特异性酶-DNA相互作用的过程.

主要方法:

  • 使用X射线晶体学来确定与DNA复合的菌体T4DNA-腺氨基甲基转移酶 (T4Dam) 的结构.
  • 对大肠杆菌DNA甲基转移酶 (EcoDam) 进行了变异性研究,以分析残留物替代的影响.
  • 生物化学试验被用来评估野生类型和突变酶的甲基化活性和特异性.

主要成果:

  • 确定了T4Dam在三元复合体中的三个晶体结构,其中包括DNA和甲基捐赠器模拟物.
  • 该研究确定了两种不同类型的蛋白质-DNA相互作用:歧视性和反歧视性接触.

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Genome-wide Mapping of Protein-DNA Interactions with ChEC-seq in Saccharomyces cerevisiae
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Genome-wide Mapping of Protein-DNA Interactions with ChEC-seq in Saccharomyces cerevisiae

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Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors
06:07

Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors

Published on: August 5, 2022

相关实验视频

Last Updated: May 12, 2026

DamID-seq: Genome-wide Mapping of Protein-DNA Interactions by High Throughput Sequencing of Adenine-methylated DNA Fragments
09:14

DamID-seq: Genome-wide Mapping of Protein-DNA Interactions by High Throughput Sequencing of Adenine-methylated DNA Fragments

Published on: January 28, 2016

Genome-wide Mapping of Protein-DNA Interactions with ChEC-seq in Saccharomyces cerevisiae
10:43

Genome-wide Mapping of Protein-DNA Interactions with ChEC-seq in Saccharomyces cerevisiae

Published on: June 3, 2017

Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors
06:07

Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors

Published on: August 5, 2022

  • 在EcoDam中对应于T4Dam中特异性决定残留的突变改变了DNA甲基化模式.
  • 结论:

    • 歧视性接触稳定了过渡状态,并增强了相关DNA位点的甲基化.
    • 反歧视性接触减少了非同源地点的甲基化,有助于序列特异性.
    • 这些发现说明了酶-DNA相互作用的逐步过渡,从非特异性接触到特异性接触,表明它们形成的时间顺序.