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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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Related Experiment Video

Updated: Nov 21, 2025

Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51
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RADX controls RAD51 filament dynamics to regulate replication fork stability.

Madison B Adolph1, Taha M Mohamed1, Swati Balakrishnan2

  • 1Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA.

Molecular Cell
|January 16, 2021
PubMed
Summary
This summary is machine-generated.

RADX protein regulates DNA replication by destabilizing RAD51 nucleofilaments, preventing genome instability. It opposes BRCA2

Keywords:
DNA curtainDNA damage responseDNA repairdouble-strand breakelectron microscopyfork reversalreplication stress

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • RAD51 recombinase is crucial for DNA double-strand break repair and replication fork stability.
  • Dysregulation of RAD51 activity can lead to genome instability.
  • RADX is a single-strand DNA (ssDNA) binding protein involved in DNA replication regulation.

Purpose of the Study:

  • To elucidate the mechanism of action of RADX in regulating DNA replication.
  • To define how RADX interacts with and affects RAD51 nucleoprotein filaments.

Main Methods:

  • Biochemical assays to assess RAD51 strand exchange and D-loop formation.
  • Analysis of RADX interaction with RAD51, including ATP hydrolysis.
  • Assessment of RADX's role in maintaining replication fork stability in vitro and in vivo.

Main Results:

  • RADX directly inhibits RAD51 strand exchange and D-loop formation.
  • RADX selectively binds to ATP-bound RAD51, promoting ATP hydrolysis and destabilizing RAD51 nucleofilaments.
  • RADX's ssDNA binding and RAD51 interaction are essential for maintaining replication fork elongation and stability.
  • BRCA2 can counteract RADX-mediated inhibition of RAD51.

Conclusions:

  • RADX acts as a negative regulator of RAD51 nucleofilament stability.
  • RADX functions antagonistically to BRCA2 in controlling RAD51 activity during DNA replication.
  • Proper regulation of RAD51 by RADX is vital for preventing genome instability.