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Related Concept Videos

The DNA Replication Fork01:02

The DNA Replication Fork

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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...
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S-Cdk Initiates DNA Replication02:38

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The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
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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,...
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Chromosome Replication02:31

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Before a cell can divide, it must accurately replicate all of its chromosomes, including the DNA and its associated histone and non-histone proteins.  This process begins at numerous origins of replication during the S phase of the cell cycle in each of a cell’s chromosomes simultaneously. Certain nucleotides can act as origins of replication, but these sequences are not well defined - especially in complex, multi-cellular, eukaryotic species. The length of DNA that spans an origin...
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Replication in Prokaryotes01:32

Replication in Prokaryotes

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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.
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Replication in Eukaryotes01:29

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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.
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Related Experiment Video

Updated: Sep 14, 2025

Genome-wide Determination of Mammalian Replication Timing by DNA Content Measurement
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Genome-wide Determination of Mammalian Replication Timing by DNA Content Measurement

Published on: January 19, 2017

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DNA replication initiation timing is important for maintaining genome integrity.

Tristan T Reed1, Abigail H Kendal1, Katherine J Wozniak1

  • 1Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan, USA.

Journal of Bacteriology
|July 21, 2025
PubMed
Summary
This summary is machine-generated.

Regulating DNA replication initiation is crucial for genome stability. Both under-initiation and over-initiation of DNA replication in Bacillus subtilis increase sensitivity to DNA damage, impacting repair efficiency.

Keywords:
Bacillus subtilisCcrZDNA replicationDnaARecA

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • DNA replication initiation is tightly regulated by positive and negative factors.
  • The impact of altered replication initiation frequency on bacterial genome stability is not well understood.
  • YabA acts as a negative regulator, while CcrZ acts as a positive regulator of replication initiation in Bacillus subtilis.

Purpose of the Study:

  • To investigate the consequences of DNA replication under-initiation and over-initiation on genome stability in Bacillus subtilis.
  • To determine the relationship between replication initiation frequency and sensitivity to genotoxic stress.
  • To elucidate the mechanisms underlying genome instability caused by altered replication initiation.

Main Methods:

  • Measuring the origin-to-terminus ratio as a proxy for replication initiation activity.
  • Analyzing RecA-GFP foci formation to assess replication fork stress.
  • Evaluating sensitivity to mitomycin C to determine genotoxic stress response.

Main Results:

  • Deletion of ccrZ or specific ccrZ alleles led to under-initiation of DNA replication.
  • Ablation of yabA or CcrZ overproduction resulted in over-initiation of replication.
  • Both under-initiating and over-initiating cells exhibited increased sensitivity to mitomycin C, indicating compromised DNA repair capacity.

Conclusions:

  • Tight regulation of DNA replication initiation is essential for maintaining genome integrity in bacteria.
  • Under-initiation leads to sensitivity via asynchronous replication and inefficient homologous recombination.
  • Over-initiation causes replication fork stress, exacerbating DNA damage sensitivity.