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Related Concept Videos

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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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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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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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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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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Related Experiment Video

Updated: Jun 28, 2025

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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Replisome Proximal Protein Associations and Dynamic Proteomic Changes at Stalled Replication Forks.

Carla-Marie Jurkovic1, Jennifer Raisch1, Stephanie Tran2

  • 1Faculty of Medicine and Health Sciences, Department of Immunology and Cell Biology, Université de Sherbrooke, Sherbrooke, Québec, Canada.

Molecular & Cellular Proteomics : MCP
|April 14, 2024
PubMed
Summary

Researchers mapped protein interactions during DNA replication and repair. Genotoxic stress dynamically reorganizes the replisome, revealing new proteins involved in DNA replication arrest.

Keywords:
BioIDDNA repairDNA replicationbiotinylationhydroxyureainteractomeproteomicsreplication fork

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • DNA replication is crucial for cell division, ensuring genetic information transfer.
  • Replication forks require coordinated protein assembly and are vulnerable to genotoxic stressors.
  • Understanding protein interactions is key to deciphering DNA replication and repair mechanisms.

Purpose of the Study:

  • To identify protein associations within core replication machinery.
  • To investigate how genotoxic stress impacts these interactions.
  • To uncover novel proteins involved in DNA replication stress response.

Main Methods:

  • Proximity-dependent biotin identification was employed to map protein associations.
  • The study focused on four core replication components: helicase, DNA polymerases, replication protein A, and histone chaperones.
  • The impact of hydroxyurea, a genotoxic stressor, on protein interactions was analyzed.

Main Results:

  • A network of 108 nuclear proteins associated with core replication components was identified.
  • Hydroxyurea treatment modulated these interactions, with 45 proteins enriched and 63 depleted.
  • Hydroxyurea also caused redistribution of 11 interactors, indicating replisome reorganization under stress.
  • Several poorly characterized proteins were identified as potential new players in DNA replication stress response.

Conclusions:

  • The study provides a comprehensive proteomic framework for understanding cellular responses to DNA replication obstacles.
  • It reveals the dynamic nature of the replisome under genotoxic stress.
  • New putative proteins involved in DNA replication arrest have been uncovered.