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4D Visualization of replication foci in mammalian cells corresponding to individual replicons

Author

Listed:
  • V. O. Chagin

    (Technische Universitaet Darmstadt
    Laboratory of chromosome stability, Institute of Cytology)

  • C. S. Casas-Delucchi

    (Technische Universitaet Darmstadt
    Cancer Research UK London Research Institute, Clare Hall Laboratories)

  • M. Reinhart

    (Technische Universitaet Darmstadt)

  • L. Schermelleh

    (Micron Advanced Bioimaging Unit, University of Oxford
    Ludwig Maximilians University Munich)

  • Y. Markaki

    (Ludwig Maximilians University Munich)

  • A. Maiser

    (Ludwig Maximilians University Munich)

  • J. J. Bolius

    (Technische Universitaet Darmstadt)

  • A. Bensimon

    (Genomic Vision)

  • M. Fillies

    (Max Delbrueck Center for Molecular Medicine
    Charité—Universitaetsmedizin)

  • P. Domaing

    (Max Delbrueck Center for Molecular Medicine)

  • Y. M. Rozanov

    (Laboratory of chromosome stability, Institute of Cytology)

  • H. Leonhardt

    (Ludwig Maximilians University Munich)

  • M. C. Cardoso

    (Technische Universitaet Darmstadt)

Abstract

Since the pioneering proposal of the replicon model of DNA replication 50 years ago, the predicted replicons have not been identified and quantified at the cellular level. Here, we combine conventional and super-resolution microscopy of replication sites in live and fixed cells with computational image analysis. We complement these data with genome size measurements, comprehensive analysis of S-phase dynamics and quantification of replication fork speed and replicon size in human and mouse cells. These multidimensional analyses demonstrate that replication foci (RFi) in three-dimensional (3D) preserved somatic mammalian cells can be optically resolved down to single replicons throughout S-phase. This challenges the conventional interpretation of nuclear RFi as replication factories, that is, the complex entities that process multiple clustered replicons. Accordingly, 3D genome organization and duplication can be now followed within the chromatin context at the level of individual replicons.

Suggested Citation

  • V. O. Chagin & C. S. Casas-Delucchi & M. Reinhart & L. Schermelleh & Y. Markaki & A. Maiser & J. J. Bolius & A. Bensimon & M. Fillies & P. Domaing & Y. M. Rozanov & H. Leonhardt & M. C. Cardoso, 2016. "4D Visualization of replication foci in mammalian cells corresponding to individual replicons," Nature Communications, Nature, vol. 7(1), pages 1-12, September.
    • Handle: RePEc:nat:natcom:v:7:y:2016:i:1:d:10.1038_ncomms11231
    DOI: 10.1038/ncomms11231
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    Cited by:

    1. Eri Koyanagi & Yoko Kakimoto & Tamiko Minamisawa & Fumiya Yoshifuji & Toyoaki Natsume & Atsushi Higashitani & Tomoo Ogi & Antony M. Carr & Masato T. Kanemaki & Yasukazu Daigaku, 2022. "Global landscape of replicative DNA polymerase usage in the human genome," Nature Communications, Nature, vol. 13(1), pages 1-18, December.
    2. María Arroyo & Florian D. Hastert & Andreas Zhadan & Florian Schelter & Susanne Zimbelmann & Cathia Rausch & Anne K. Ludwig & Thomas Carell & M. Cristina Cardoso, 2022. "Isoform-specific and ubiquitination dependent recruitment of Tet1 to replicating heterochromatin modulates methylcytosine oxidation," Nature Communications, Nature, vol. 13(1), pages 1-28, December.

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