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Related Concept Videos

Homologous Recombination02:31

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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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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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In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
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In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
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Visualization of DNA Repair Proteins Interaction by Immunofluorescence
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Rad51 determines pathway usage in post-replication repair.

Damon Meyer1,2, Steven K Gore1, Jie Liu1

  • 1Department of Microbiology & Molecular Genetics, University of California, Davis, CA, USA.

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|January 10, 2026
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This summary is machine-generated.

Mutations in Rad51 protein alter DNA binding, shifting post-replication repair from recombination to translesion synthesis. This highlights Rad51

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Area of Science:

  • Molecular Biology
  • Genetics
  • DNA Repair Mechanisms

Background:

  • Stalled replication forks require post-replication repair via homologous recombination, fork regression, or translesion DNA synthesis.
  • The regulation of pathway choice in post-replication repair remains incompletely understood.
  • Rad51 protein is crucial for genomic stability, participating in recombination and protecting stalled forks.

Purpose of the Study:

  • To investigate the role of Rad51 protein in regulating pathway usage during post-replication repair.
  • To identify specific Rad51 mutations that alter its function in stalled replication fork processing.
  • To elucidate the mechanism by which Rad51 controls the choice between recombination and alternative repair pathways.

Main Methods:

  • Isolation and characterization of Rad51 mutations in Saccharomyces cerevisiae.
  • In vitro biochemical assays to assess Rad51 recombination activity and DNA binding profiles.
  • In vivo studies to evaluate Rad51 recruitment to stalled replication forks.
  • In vitro assays examining the protection of double-stranded DNA (dsDNA) from degradation by nucleases.

Main Results:

  • Identified Rad51 mutations (Rad51-E135D, Rad51-K305N) that shift repair from recombination to translesion synthesis.
  • Mutant Rad51 proteins exhibited near-normal in vitro recombination but altered dsDNA binding.
  • These mutants displayed defects in Rad51 recruitment to stalled forks in vivo.
  • Mutant Rad51 proteins showed impaired protection of dsDNA from degradation by Dna2-Sgs1 and Exo1 in vitro.

Conclusions:

  • Rad51 binding to duplex DNA is essential for controlling pathway selection at stalled replication forks.
  • Specific Rad51 mutations disrupt its function in protecting stalled forks, leading to altered repair pathway usage.
  • Understanding Rad51's DNA binding properties provides insights into maintaining genomic stability.