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Related Concept Videos

Replication in Eukaryotes02:31

Replication in Eukaryotes

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Overview
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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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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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The DNA Replication Fork01:02

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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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Related Experiment Video

Updated: Feb 27, 2026

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

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Proteomic Analyses of the Eukaryotic Replication Machinery.

David Cortez1

  • 1Vanderbilt University School of Medicine, Nashville, TN, United States.

Methods in Enzymology
|June 25, 2017
PubMed
Summary

New proteomic tools like isolation of proteins on nascent DNA (iPOND) and nascent chromatin capture (NCC) help identify proteins involved in DNA replication. These methods reveal insights into maintaining genome and chromatin integrity during replication stress.

Keywords:
ChromatinDNA replicationMass spectrometryNascent chromatin captureProteomicsReplication stressReplisomeSILACiPOND

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Area of Science:

  • Molecular Biology
  • Proteomics
  • Cell Biology

Background:

  • DNA replication requires hundreds of proteins for accurate duplication each cell cycle.
  • Accessory proteins manage replication stress, error correction, and chromatin packaging.
  • Understanding these protein complexes is crucial for genome integrity.

Purpose of the Study:

  • To introduce novel proteomic tools for studying DNA replication.
  • To examine the application of these tools in defining the replication fork proteome.
  • To highlight discoveries in replication, chromatin maturation, and stress response.

Main Methods:

  • Utilized proteomic tools such as isolation of proteins on nascent DNA (iPOND).
  • Employed nascent chromatin capture (NCC) for protein identification and quantification.
  • Applied these methods to purify and analyze replication and chromatin machineries.

Main Results:

  • Discovered new proteins involved in DNA replication and chromatin maturation.
  • Gained insights into the dynamic regulatory processes ensuring genome integrity.
  • Defined key components of the replication fork proteome.

Conclusions:

  • Proteomic advancements have significantly enhanced the study of DNA replication.
  • iPOND and NCC are powerful techniques for dissecting complex cellular processes.
  • These tools are vital for understanding genome stability and response to replication stress.