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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...
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...
Replication in Prokaryotes01:32

Replication in Prokaryotes

DNA replication has three main steps: initiation, elongation, and termination. Replication in prokaryotes begins when initiator proteins bind to the single origin of replication (ori) on the cell's circular chromosome. Replication then proceeds around the entire circle of the chromosome in each direction from the two replication forks, resulting in two DNA molecules.
Many Proteins Work Together to Replicate the Chromosome
Replication is coordinated and carried out by a host of specialized...
Replication in Prokaryotes02:35

Replication in Prokaryotes

Overview

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Related Experiment Video

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Visualization of DNA Repair Proteins Interaction by Immunofluorescence
07:55

Visualization of DNA Repair Proteins Interaction by Immunofluorescence

Published on: June 26, 2020

Migrating bubble during break-induced replication drives conservative DNA synthesis.

Natalie Saini1, Sreejith Ramakrishnan, Rajula Elango

  • 1School of Biology and Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.

Nature
|September 13, 2013
PubMed
Summary

Break-induced replication (BIR), a DNA repair pathway, causes genetic instability. Our study reveals BIR uses a unique replication fork, not standard semiconservative replication, leading to increased mutations and potential cancer development.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Chromosomal double-strand breaks (DSBs) require repair for genomic integrity.
  • DSB repair pathways, like break-induced replication (BIR), can paradoxically cause genetic instability, mutations, and chromosomal rearrangements, driving carcinogenesis.
  • BIR is known to promote genetic instability, including increased mutation rates, loss of heterozygosity, and copy number variations.

Purpose of the Study:

  • To investigate the mechanism of DNA replication during break-induced replication (BIR).
  • To determine if BIR proceeds via semiconservative replication like normal S-phase replication.
  • To elucidate the cause of the high mutation rate associated with BIR.

Main Methods:

  • Utilized budding yeast as a model organism.
  • Analyzed the replication fork structure during BIR.
  • Investigated the role of Pif1 helicase in BIR-associated mutagenesis.

Main Results:

  • Demonstrated that BIR replication in budding yeast proceeds via an unusual bubble-like replication fork.
  • Showed that this atypical replication mechanism results in conservative inheritance of newly synthesized DNA.
  • Provided evidence that the Pif1 helicase is critical for this mode of replication and the associated increase in mutations.

Conclusions:

  • BIR replication differs significantly from S-phase replication, employing a distinct, unstable fork structure.
  • This unique replication mechanism is responsible for the elevated mutation rates observed during BIR.
  • BIR-driven synthesis represents a potent source of genetic instability in eukaryotes, potentially contributing to cancer initiation.