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Replication in Eukaryotes01:29

Replication in Eukaryotes

14.8K
In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
Many Proteins Orchestrate Replication at the Origin
Eukaryotic replication follows many of the same...
14.8K
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

5.9K
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,...
5.9K
Replication in Prokaryotes01:32

Replication in Prokaryotes

25.4K
DNA replication has three main steps: initiation, elongation, and termination. Replication in prokaryotes begins when initiator proteins bind to the single origin of replication (ori) on the cell's circular chromosome. Replication then proceeds around the entire circle of the chromosome in each direction from the two replication forks, resulting in two DNA molecules.
Many Proteins Work Together to Replicate the Chromosome
Replication is coordinated and carried out by a host of specialized...
25.4K
The DNA Replication Fork01:02

The DNA Replication Fork

37.0K
An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication...
37.0K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.2K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
6.2K
Chromosome Structure02:40

Chromosome Structure

23.7K
A functional eukaryotic chromosome must contain three elements: a centromere, telomeres, and numerous origins of replication.
The centromere is a DNA sequence that links sister chromatids. This is also where kinetochores, protein complexes to which spindle microtubules attach, are constructed after the chromosome is replicated. The kinetochores allow the spindle microtubules to move the chromosomes within the cell during cell division.
Telomeres consist of non-coding repetitive nucleotide...
23.7K

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相关实验视频

Updated: Sep 19, 2025

Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level
10:11

Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level

Published on: July 26, 2024

1.2K

一个Orc6 tether在复制起源许可期间调解ORC绑定站点切换.

David Driscoll1, Larry J Friedman2, Jeff Gelles2

  • 1Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

bioRxiv : the preprint server for biology
|June 4, 2025
PubMed
概括

原始识别复合体 (ORC) 使用Orc6在DNA复制原始授权过程中将Mcm2-7螺旋酶结合起来. 这种绑定防止了ORC解离,并解释了一个ORC如何加载两个螺旋酶.

科学领域:

  • 分子生物学分子生物学
  • 细胞生物学 细胞生物学
  • 生物化学 生化学

背景情况:

  • 启动DNA复制是细胞分裂的一个基本过程.
  • 原产地识别复合体 (ORC) 和Mcm2-7酶对于许可复制原产地至关重要.
  • 一个关键的步骤是ORC将两个Mcm2-7六合体加载到DNA上.

研究的目的:

  • 为了阐明单个ORC如何加载两个Mcm2-7螺旋酶的分子机制.
  • 为了研究Orc6在螺旋酶加载过程中的作用.
  • 了解CDK酸化如何影响原产地许可期间的ORC功能.

主要方法:

  • 采用了单分子福斯特共振能量转移 (smFRET) 试验.
  • 对Orc6进行了突变分析.
  • 生物化学实验是为了研究蛋白相互作用而进行的.

主要成果:

  • 在关键结合位过渡过程中,Orc6的N终端半端充当带,将ORC连接到Mcm2-7螺旋酶.
  • 这种Orc6介导的绑定防止ORC与DNA和螺旋酶脱离.
  • 对ORC的CDK酸化破坏了这种结合相互作用,抑制了Mcm2-7双六合体的形成.

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Visualizing the Interaction Between the Qdot-labeled Protein and Site-specifically Modified λ DNA at the Single Molecule Level
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Visualizing the Interaction Between the Qdot-labeled Protein and Site-specifically Modified λ DNA at the Single Molecule Level

Published on: July 17, 2018

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DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation
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DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation

Published on: December 29, 2021

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相关实验视频

Last Updated: Sep 19, 2025

Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level
10:11

Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level

Published on: July 26, 2024

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Visualizing the Interaction Between the Qdot-labeled Protein and Site-specifically Modified λ DNA at the Single Molecule Level
08:56

Visualizing the Interaction Between the Qdot-labeled Protein and Site-specifically Modified λ DNA at the Single Molecule Level

Published on: July 17, 2018

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DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation
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DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation

Published on: December 29, 2021

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  • 特定的Orc6链接器突变支持初始复合体形成,但阻碍了第二个Mcm2-7六合体的稳定招募.
  • 结论:

    • Orc6在促进单个ORC装载两个Mcm2-7螺旋中发挥着关键作用.
    • Orc6连接机制解释了原产地许可证的效率和忠实性.
    • Orc6是一个关键调节器,参与多个阶段的起源激活,链接DNA结合,酶负载和细胞周期控制.