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Related Concept Videos

Homologous Recombination02:31

Homologous Recombination

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...
Homologous Recombination02:31

Homologous Recombination

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...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

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, a...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
Crossing Over01:30

Crossing Over

Crossing over is the exchange of genetic information between homologous chromosomes during prophase I of meiosis I. Genetic recombination gives rise to allelic diversity in the newly formed daughter cells. In humans, crossing over produces genetically distinct haploid egg and sperm cells that undergo fertilization to produce unique offspring. Before cell division starts, the germ cell’s chromosome(s) undergo duplication in the S phase of the cell cycle. As the cells enter prophase I, duplicated...

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Related Experiment Video

Updated: Jun 12, 2026

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae
07:55

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

RPA-ssDNA co-phase separation facilitates RAD51 enrichment during homologous recombination.

Yanan Li1,2, Yi Zhao2, Yinghong Chen3,4

  • 1College of Life Sciences, Xinyang Normal University, Xinyang, Henan 464000, China.

Nucleic Acids Research
|June 11, 2026
PubMed
Summary
This summary is machine-generated.

Replication protein A (RPA) forms co-condensates with DNA, enriching RAD51 for homologous recombination (HR) DNA repair. RPA70N domain is key for this process, impacting genome stability.

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Visualization of DNA Repair Proteins Interaction by Immunofluorescence

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Related Experiment Videos

Last Updated: Jun 12, 2026

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae
07:55

Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae

Published on: September 11, 2022

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51
06:24

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51

Published on: February 13, 2019

Visualization of DNA Repair Proteins Interaction by Immunofluorescence
07:55

Visualization of DNA Repair Proteins Interaction by Immunofluorescence

Published on: June 26, 2020

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Homologous recombination (HR) maintains genome integrity by repairing DNA double-strand breaks (DSB).
  • Replication protein A (RPA) coats single-stranded DNA (ssDNA) during HR, but how RAD51 identifies RPA-coated ssDNA for replacement is unclear.

Purpose of the Study:

  • To investigate the mechanism by which RAD51 identifies RPA-coated ssDNA during homologous recombination.
  • To elucidate the role of RPA in facilitating RAD51 loading onto ssDNA for DSB repair.

Main Methods:

  • Developed a novel method for generating ssDNA for single-molecule measurements.
  • Utilized in vitro and in vivo assays to study RPA-ssDNA interactions and RAD51 enrichment.
  • Investigated the role of the RPA70 N-terminal domain (RPA70N) in RPA phase separation and RAD51 recruitment.

Main Results:

  • Human RPA undergoes phase separation, forming co-condensates with ssDNA, mediated by the RPA70N domain.
  • These RPA-ssDNA co-condensates are stable nucleoprotein assemblies that enrich RAD51 locally.
  • Deletion or sequestration of RPA70N disrupts RPA phase separation, reduces RAD51 enrichment, and impairs DSB repair.

Conclusions:

  • RPA phase separation and co-condensation with ssDNA are critical for RAD51 enrichment during HR.
  • The RPA70N domain is a key determinant for RPA-ssDNA co-condensate formation and subsequent RAD51 recruitment.
  • This mechanism highlights a novel role for RPA in facilitating early-stage HR-directed DSB repair.