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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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Direct Restart of a Replication Fork Stalled by a Head-On RNA Polymerase
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Terminating the replication helicase.

Vincent Gaggioli1, Philip Zegerman1

  • 1Wellcome Trust/Cancer Research UK Gurdon Institute and the Department of Biochemistry, The Henry Wellcome Building of Cancer and Developmental Biology, University of Cambridge, Cambridge CB2 1QN, UK.

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Summary
This summary is machine-generated.

The cell cycle requires resetting before each new phase. A conserved pathway involving ubiquitin and CDC48 (p97) removes replication machinery from DNA, returning chromosomes to an unreplicated state.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • The cell cycle involves distinct phases that must be properly regulated.
  • Resetting cellular machinery is crucial for accurate progression through the cell cycle.
  • Eukaryotic chromosome replication must be followed by a reset to the unreplicated state.

Purpose of the Study:

  • To elucidate the mechanism by which replication machinery is removed from DNA after replication.
  • To identify the key molecular players involved in resetting chromosomes to an unreplicated state.
  • To understand how eukaryotic chromosomes are prepared for subsequent cell cycle events.

Main Methods:

  • Investigated the role of ubiquitination in regulating cell cycle progression.
  • Utilized biochemical assays to study protein-protein interactions.
  • Employed genetic approaches to analyze the function of CDC48 (p97) in DNA replication and cell cycle control.

Main Results:

  • Identified a conserved ubiquitin- and CDC48 (p97)-dependent pathway responsible for replication machinery removal.
  • Demonstrated that this pathway actively disassembles the replication complex from fully replicated DNA.
  • Showed that the removal of replication machinery is essential for returning chromosomes to an unreplicated state.

Conclusions:

  • The study reveals a fundamental mechanism for cell cycle resetting.
  • Ubiquitin and CDC48 (p97) play a critical role in ensuring proper chromosome status for the next cell cycle.
  • This conserved pathway is key to maintaining genomic integrity and cell cycle fidelity in eukaryotes.