1. Chromosomes and Gene Expression

TRAIP drives replisome disassembly and mitotic DNA repair synthesis at sites of incomplete DNA replication

  1. Remi Sonneville
  2. Rahul Bhowmick
  3. Saskia Hoffmann
  4. Niels mailand  Is a corresponding author
  5. Ian D Hickson  Is a corresponding author
  6. Karim Labib  Is a corresponding author
  1. University of Dundee, United Kingdom
  2. University of Copenhagen, Denmark
https://doi.org/10.7554/eLife.48686
Share this article
Cite this article
  1. Remi Sonneville
  2. Rahul Bhowmick
  3. Saskia Hoffmann
  4. Niels mailand
  5. Ian D Hickson
  6. Karim Labib
(2019)
TRAIP drives replisome disassembly and mitotic DNA repair synthesis at sites of incomplete DNA replication
eLife 8:e48686.
https://doi.org/10.7554/eLife.48686
  2. Download BibTeX
  3. Download .RIS

Abstract

The faithful segregation of eukaryotic chromosomes in mitosis requires that the genome be duplicated completely prior to anaphase. However, cells with large genomes sometimes fail to complete replication during interphase and instead enter mitosis with regions of incompletely replicated DNA. These regions are processed in early mitosis via a process known as mitotic DNA repair synthesis (MiDAS), but little is known about how cells switch from conventional DNA replication to MiDAS. Using the early embryo of the nematode Caenorhabditis elegans as a model system, we show that the TRAIP ubiquitin ligase drives replisome disassembly in response to incomplete DNA replication, thereby providing access to replication forks for other factors. Moreover, TRAIP is essential for MiDAS in human cells, and is important in both systems to prevent mitotic segregation errors. Our data indicate that TRAIP is a master regulator of the processing of incomplete DNA replication during mitosis in metazoa.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Remi Sonneville

    MRC Protein Phosphorylation and Ubiquitylation Unit, University of Dundee, Dundee, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.

  2. Rahul Bhowmick

    Center for Chromosome Stability, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    The authors declare that no competing interests exist.

  3. Saskia Hoffmann

    Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    The authors declare that no competing interests exist.

  4. Niels mailand

    Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
    For correspondence
    niels.mailand@cpr.ku.dk
    Competing interests
    The authors declare that no competing interests exist.

  5. Ian D Hickson

    Center for Chromosome Stability, University of Copenhagen, Copenhagen, Denmark
    For correspondence
    iandh@sund.ku.dk
    Competing interests
    The authors declare that no competing interests exist.

  6. Karim Labib

    MRC Protein Phosphorylation and Ubiquitylation Unit, University of Dundee, Dundee, United Kingdom
    For correspondence
    kpmlabib@dundee.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8861-379X

Funding

Medical Research Council (MC_UU_12016/13)

  • Remi Sonneville
  • Karim Labib

Cancer Research UK (C578/A24558)

  • Remi Sonneville
  • Karim Labib

Wellcome (102943/Z/13/Z)

  • Karim Labib

Danish National Research Foundation (DNRF115)

  • Ian D Hickson

Danish Medical Research Council (DFF-4004-00155B)

  • Rahul Bhowmick

H2020 European Research Council (616236)

  • Niels mailand

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

Reviewing Editor

  1. Andrés Aguilera, CABIMER, Universidad de Sevilla, Spain

Publication history

  1. Received: May 22, 2019
  2. Accepted: September 20, 2019
  3. Accepted Manuscript published: September 23, 2019 (version 1)
  4. Version of Record published: October 1, 2019 (version 2)

Copyright

© 2019, Sonneville 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.

Metrics

  • 2,522
    Page views
  • 539
    Downloads
  • 41
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, PubMed Central.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Remi Sonneville
  2. Rahul Bhowmick
  3. Saskia Hoffmann
  4. Niels mailand
  5. Ian D Hickson
  6. Karim Labib
(2019)
TRAIP drives replisome disassembly and mitotic DNA repair synthesis at sites of incomplete DNA replication
eLife 8:e48686.
https://doi.org/10.7554/eLife.48686

Categories and tags

Research organisms

Further reading

    1. Cell Biology
    2. Chromosomes and Gene Expression

    Overcoming the cytoplasmic retention of GDOWN1 modulates global transcription and facilitates stress adaptation

    Zhanwu Zhu, Jingjing Liu ... Bo Cheng
    Research Article

    Dynamic regulation of transcription is crucial for the cellular responses to various environmental or developmental cues. Gdown1 is a ubiquitously expressed, RNA polymerase II (Pol II) interacting protein, essential for the embryonic development of metazoan. It tightly binds Pol II in vitro and competitively blocks the binding of TFIIF and possibly other transcriptional regulatory factors, yet its cellular functions and regulatory circuits remain unclear. Here, we show that human GDOWN1 strictly localizes in the cytoplasm of various types of somatic cells and exhibits a potent resistance to the imposed driving force for its nuclear localization. Combined with the genetic and microscope-based approaches, two types of the functionally coupled and evolutionally conserved localization regulatory motifs are identified, including the CRM1-dependent nucleus export signal (NES) and a novel Cytoplasmic Anchoring Signal (CAS) that mediates its retention outside of the nuclear pore complexes (NPC). Mutagenesis of CAS alleviates GDOWN1’s cytoplasmic retention, thus unlocks its nucleocytoplasmic shuttling properties, and the increased nuclear import and accumulation of GDOWN1 results in a drastic reduction of both Pol II and its associated global transcription levels. Importantly, the nuclear translocation of GDOWN1 occurs in response to the oxidative stresses, and the ablation of GDOWN1 significantly weakens the cellular tolerance. Collectively, our work uncovers the molecular basis of GDOWN1’s subcellular localization and a novel cellular strategy of modulating global transcription and stress-adaptation via controlling the nuclear translocation of GDOWN1.

    1. Chromosomes and Gene Expression

    Cancer: Examining the cooperation between extrachromosomal DNA circles

    Jiahui Zhang, Cheng-Zhong Zhang
    Insight

    In a departure from previous findings, new results suggest that free-floating pieces of DNA which carry additional copies of cancer-driving genes do not tend to cluster or have increased transcription.

    1. Chromosomes and Gene Expression

    Oncogene expression from extrachromosomal DNA is driven by copy number amplification and does not require spatial clustering in glioblastoma stem cells

    Karin Purshouse, Elias T Friman ... Wendy A Bickmore
    Research Article

    Extrachromosomal DNA (ecDNA) are frequently observed in human cancers and are responsible for high levels of oncogene expression. In glioblastoma (GBM), ecDNA copy number correlates with poor prognosis. It is hypothesized that their copy number, size, and chromatin accessibility facilitate clustering of ecDNA and colocalization with transcriptional hubs, and that this underpins their elevated transcriptional activity. Here, we use super-resolution imaging and quantitative image analysis to evaluate GBM stem cells harbouring distinct ecDNA species (EGFR, CDK4, PDGFRA). We find no evidence that ecDNA routinely cluster with one another or closely interact with transcriptional hubs. Cells with EGFR-containing ecDNA have increased EGFR transcriptional output, but transcription per gene copy is similar in ecDNA compared to the endogenous chromosomal locus. These data suggest that it is the increased copy number of oncogene-harbouring ecDNA that primarily drives high levels of oncogene transcription, rather than specific interactions of ecDNA with each other or with high concentrations of the transcriptional machinery.