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Related Concept Videos

S-Cdk Initiates DNA Replication02:38

S-Cdk Initiates DNA Replication

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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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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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Lagging Strand Synthesis01:59

Lagging Strand Synthesis

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During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
There are several major differences between synthesis of the leading strand and synthesis of the lagging strand. 1) Leading strand synthesis happens in the direction of replication fork opening, whereas lagging strand synthesis happens in the...
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Restarting Stalled Replication Forks02:37

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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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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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The Replisome03:01

The Replisome

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DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
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Chromosome Replicating Timing Combined with Fluorescent In situ Hybridization
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Kronos scRT: a uniform framework for single-cell replication timing analysis.

Stefano Gnan1, Joseph M Josephides1, Xia Wu1,2

  • 1Institut Curie, PSL Research University, CNRS UMR3244, Dynamics of Genetic Information, Sorbonne Université, 75005, Paris, France.

Nature Communications
|April 28, 2022
PubMed
Summary
This summary is machine-generated.

We developed Kronos scRT, a new software for single-cell Replication Timing (scRT) analysis. This tool enables large-scale studies of genome replication timing in thousands of cells without cell sorting.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Computational Biology

Background:

  • Mammalian genome replication timing is cell type-specific and linked to transcription and chromatin organization.
  • Existing single-cell replication studies are limited by low cell numbers (<100) due to individual cell processing and sorting requirements.

Purpose of the Study:

  • To develop Kronos scRT, a novel software tool for single-cell Replication Timing (scRT) analysis.
  • To enable high-throughput scRT analysis from large, asynchronous cell populations without cell sorting.

Main Methods:

  • Development of Kronos scRT software for single-cell Replication Timing analysis.
  • Application of Kronos scRT to analyze published and newly generated droplet-based single-cell whole-genome sequencing data.
  • Analysis of thousands of cells from mouse and human cell lines.

Main Results:

  • Kronos scRT facilitates large-scale scRT analysis from thousands of cells.
  • Genomic regions generally replicate around their population average Replication Timing (RT).
  • Replication timing exhibits stochasticity across the S phase in single cells.

Conclusions:

  • Kronos scRT enables fast and comprehensive investigation of Replication Timing (RT) programs at single-cell resolution.
  • The tool is applicable to both homogeneous and heterogeneous cell populations.
  • Demonstrates stochasticity in genome replication timing at the single-cell level.