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Related Concept Videos

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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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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 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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Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
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Priming for tolerance and cohesion at replication forks.

Dana Branzei1, Barnabas Szakal1

  • 1a IFOM, the FIRC Institute of Molecular Oncology , Milan , Italy.

Nucleus (Austin, Tex.)
|February 19, 2016
PubMed
Summary
This summary is machine-generated.

Genome duplication involves DNA damage tolerance (DDT) and chromatin changes. Our study reveals that managing single-stranded DNA at replication forks is key to chromosome structure and DDT pathway choice.

Keywords:
Chromosome replicationDNA damage tolerancefork reversal recombinationreprimingsister chromatid cohesion

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Genome duplication is intrinsically linked to DNA damage tolerance (DDT) and chromatin organization.
  • Replication fork integrity and sister chromatid cohesion (SCC) are crucial for accurate chromosome segregation.
  • Previous work identified roles for Primase subunits and Ctf4 in regulating DDT and SCC.

Purpose of the Study:

  • To investigate the relationship between replication fork dynamics, DDT, and sister chromatid cohesion.
  • To elucidate the role of single-stranded DNA management in determining replication fork structure and DDT pathway selection.
  • To understand how cohesin influences recombination-mediated DDT.

Main Methods:

  • Genetic analyses of replication mutants.
  • Electron microscopy of replication intermediates.
  • Sister chromatid tethering experiments.

Main Results:

  • Mutations in Primase or Ctf4 lead to aberrant DDT, reduced SCC, and increased fork reversal.
  • Cohesin facilitates recombination-dependent DDT.
  • Artificial sister chromatid tethering rescues cohesin but not Polα/Primase/Ctf4 mutant defects.
  • Management of single-stranded DNA near the fork is critical for chromosome and fork structure.

Conclusions:

  • Single-stranded DNA management at replication forks is a critical determinant of chromosome and replication fork structure.
  • This management influences the choice of DNA damage tolerance pathways.
  • Findings provide insights into DDT regulation and cohesion establishment during replication.