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Related Concept Videos

Replication in Eukaryotes02:31

Replication in Eukaryotes

Overview
Chromosome Structure02:40

Chromosome Structure

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...
Chromosome Replication02:31

Chromosome Replication

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 of...
Replication in Eukaryotes02:31

Replication in Eukaryotes

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Replication in Eukaryotes01:29

Replication in Eukaryotes

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...
Replication in Eukaryotes01:29

Replication in Eukaryotes

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...

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Related Experiment Video

Updated: May 28, 2026

Chromosome Replicating Timing Combined with Fluorescent In situ Hybridization
17:14

Chromosome Replicating Timing Combined with Fluorescent In situ Hybridization

Published on: December 10, 2012

Mapping replicational sites in the eucaryotic cell nucleus.

H Nakayasu1, R Berezney

  • 1Department of Biological Sciences, State University of New York, Buffalo 14260.

The Journal of Cell Biology
|January 1, 1989
PubMed
Summary
This summary is machine-generated.

Researchers mapped DNA replication sites in cell nuclei using fluorescent microscopy. They identified distinct patterns of replication granules, revealing their association with the nuclear matrix even after significant DNA removal.

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Related Experiment Videos

Last Updated: May 28, 2026

Chromosome Replicating Timing Combined with Fluorescent In situ Hybridization
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Published on: December 10, 2012

Imaging Replicative Domains in Ultrastructurally Preserved Chromatin by Electron Tomography
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Imaging Replicative Domains in Ultrastructurally Preserved Chromatin by Electron Tomography

Published on: May 20, 2022

Mapping Absolute DNA Density in Cell Nuclei using Single-molecule Localization Microscopy
10:57

Mapping Absolute DNA Density in Cell Nuclei using Single-molecule Localization Microscopy

Published on: November 11, 2025

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Understanding the spatial organization of DNA replication within the interphase nucleus is crucial for comprehending genome stability and cell cycle regulation.
  • Previous studies have suggested the existence of discrete replication sites, but their precise localization and association with nuclear structures remain areas of active investigation.

Purpose of the Study:

  • To visualize and characterize the distribution of DNA replication sites within the interphase nucleus of mammalian cells.
  • To investigate the relationship between DNA replication sites, chromatin domains, and the nuclear matrix throughout S phase.
  • To determine the structural basis for the maintenance of replication sites during DNA extraction.

Main Methods:

  • Fluorescent microscopy was employed to map DNA replication sites following the incorporation of biotinylated dUTP into permeabilized PtK-1 and 3T3 cells.
  • Antibodies against 5-bromodeoxyuridine were used to confirm replication patterns in vivo.
  • Cells were extracted with DNase I and ammonium sulfate to analyze the association of replication sites with the nuclear matrix.

Main Results:

  • Discrete DNA replication granules were observed throughout the nuclear interior and along the nuclear periphery.
  • Three distinct patterns of replication sites (Type I, II, and III) were identified, correlating with different stages of S phase (early to mid, mid to late, and late S phase, respectively).
  • Replication granules and their intranuclear arrangements were maintained with nuclear matrix structures even after removing over 90% of nuclear DNA, and the nuclear matrix itself incorporated biotinylated dUTP into replication granules.

Conclusions:

  • DNA replication occurs at discrete sites organized within the cell nucleus, exhibiting distinct spatial patterns throughout S phase.
  • These replication sites are intimately associated with the nuclear matrix, suggesting a role for the nuclear scaffold in organizing DNA replication.
  • The nuclear matrix provides a structural framework that maintains the organization of replication sites, even under conditions of extensive DNA depletion.