1. Chromosomes and Gene Expression
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Checkpoint inhibition of origin firing prevents inappropriate replication outside of S-phase

  1. Mark C Johnson
  2. Geylani Can
  3. Miguel Monteiro Santos
  4. Diana Alexander
  5. Philip Zegerman  Is a corresponding author
  1. Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, United Kingdom
Research Article
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Cite this article as: eLife 2021;10:e63589 doi: 10.7554/eLife.63589

Abstract

Checkpoints maintain the order of cell cycle events during DNA damage or incomplete replication. How the checkpoint response is tailored to different phases of the cell cycle remains poorly understood. The S-phase checkpoint for example results in the slowing of replication, which in budding yeast occurs by Rad53-dependent inhibition of the initiation factors Sld3 and Dbf4. Despite this, we show here that Rad53 phosphorylates both of these substrates throughout the cell cycle at the same sites as in S-phase, suggesting roles for this pathway beyond S-phase. Indeed, we show that Rad53-dependent inhibition of Sld3 and Dbf4 limits re-replication in G2/M, preventing gene amplification. In addition, we show that inhibition of Sld3 and Dbf4 in G1 prevents premature initiation at all origins at the G1/S transition. This study redefines the scope of the 'S-phase checkpoint' with implications for understanding checkpoint function in cancers that lack cell cycle controls.

Data availability

Sequencing data has been deposited in GEO under the accession code GSE159122 and GSE163571

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Mark C Johnson

    Biochemistry, Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Geylani Can

    Biochemistry, Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1716-7830
  3. Miguel Monteiro Santos

    Biochemistry, Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5594-2682
  4. Diana Alexander

    Biochemistry, Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7785-3170
  5. Philip Zegerman

    Biochemistry, Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, United Kingdom
    For correspondence
    paz20@cam.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5707-1083

Funding

Worlwide Cancer Research (AICR 10-0908)

  • Mark C Johnson
  • Geylani Can
  • Miguel Monteiro Santos
  • Diana Alexander
  • Philip Zegerman

Wellcome Trust (107056/Z/15/Z)

  • Mark C Johnson
  • Geylani Can
  • Miguel Monteiro Santos
  • Diana Alexander
  • Philip Zegerman

Biotechnology and Biological Sciences Research Council (BB/M011194/1)

  • Miguel Monteiro Santos

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Tim Formosa, University of Utah School of Medicine, United States

Publication history

  1. Received: September 29, 2020
  2. Accepted: January 4, 2021
  3. Accepted Manuscript published: January 5, 2021 (version 1)
  4. Version of Record published: January 13, 2021 (version 2)

Copyright

© 2021, Johnson et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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