KAT2-mediated acetylation switches the mode of PALB2 chromatin association to safeguard genome integrity

  1. Marjorie Fournier  Is a corresponding author
  2. Amélie Rodrigue
  3. Larissa Milano
  4. Jean-Yves Bleuyard
  5. Anthony M Couturier
  6. Jacob Wall
  7. Jessica Ellins
  8. Svenja Hester
  9. Stephen J Smerdon
  10. László Tora
  11. Jean-Yves Masson  Is a corresponding author
  12. Fumiko Esashi  Is a corresponding author
  1. University of Oxford, United Kingdom
  2. Laval University, Canada
  3. University of Birmingham, United Kingdom
  4. Institut de Génétique et de Biologie Moléculaire et Cellulaire, France

Abstract

The tumour suppressor PALB2 stimulates RAD51-mediated homologous recombination (HR) repair of DNA damage, whilst its steady-state association with active genes protects these loci from replication stress. Here, we report that the lysine acetyltransferases 2A and 2B (KAT2A/2B, also called GCN5/PCAF), two well-known transcriptional regulators, acetylate a cluster of seven lysine residues (7K-patch) within the PALB2 chromatin association motif (ChAM) and, in this way, regulate context-dependent PALB2 binding to chromatin. In unperturbed cells, the 7K-patch is targeted for KAT2A/2B-mediated acetylation, which in turn enhances the direct association of PALB2 with nucleosomes. Importantly, DNA damage triggers a rapid deacetylation of ChAM and increases the overall mobility of PALB2. Distinct missense mutations of the 7K-patch render the mode of PALB2 chromatin binding, making it either unstably chromatin-bound (7Q) or randomly bound with a reduced capacity for mobilisation (7R). Significantly, both of these mutations confer a deficiency in RAD51 foci formation and increase DNA damage in S phase, leading to the reduction of overall cell survival. Thus, our study reveals that acetylation of the ChAM 7K-patch acts as a molecular switch to enable dynamic PALB2 shuttling for HR repair while protecting active genes during DNA replication.

Data availability

The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (Perez-Riverol et al, 2019) partner repository with the dataset identifier PXD014678 and PXD014681. All other data generated or analysed during this study are included in the manuscript and supporting file. Raw images of western blots and DNA/protein gels are avilable through the Open Science Framework, with the following link: https://osf.io/8e9ms/?view_only=3908731b938e4751af7518744f3ff584

Article and author information

Author details

  1. Marjorie Fournier

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    For correspondence
    marjorie.fournier@path.ox.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
  2. Amélie Rodrigue

    Department of Molecular Biology, Laval University, Québec, Canada
    Competing interests
    The authors declare that no competing interests exist.
  3. Larissa Milano

    Department of Molecular Biology, Laval University, Québec, Canada
    Competing interests
    The authors declare that no competing interests exist.
  4. Jean-Yves Bleuyard

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6727-5362
  5. Anthony M Couturier

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1512-9558
  6. Jacob Wall

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  7. Jessica Ellins

    Department of Biochemistry, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  8. Svenja Hester

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  9. Stephen J Smerdon

    Institute of Cancer and Genomic Sciences, University of Birmingham, Birmingham, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  10. László Tora

    Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7398-2250
  11. Jean-Yves Masson

    Department of Molecular Biology, Laval University, Québec, Canada
    For correspondence
    Jean-Yves.Masson@crchudequebec.ulaval.ca
    Competing interests
    The authors declare that no competing interests exist.
  12. Fumiko Esashi

    Sir William Dunn School of Pathology, University of Oxford, Oxford, United Kingdom
    For correspondence
    fumiko.esashi@path.ox.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0902-2364

Funding

Wellcome Trust ((101009/Z/13/Z)

  • Fumiko Esashi

Cancer Research UK (FC001156))

  • Stephen J Smerdon

Medical Research Council (FC001156))

  • Stephen J Smerdon

Wellcome Trust (FC001156)

  • Stephen J Smerdon

H2020 European Research Council (ERC-2013-340551)

  • László Tora

Edward P Abraham Research Fund (RF 260)

  • Fumiko Esashi

Canadian Institutes of Health Research (FDN-388879)

  • Jean-Yves Masson

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

Reviewing Editor

  1. Wolf-Dietrich Heyer, University of California, Davis, United States

Publication history

  1. Preprint posted: August 15, 2019 (view preprint)
  2. Received: April 10, 2020
  3. Accepted: October 20, 2022
  4. Accepted Manuscript published: October 21, 2022 (version 1)
  5. Version of Record published: November 17, 2022 (version 2)

Copyright

© 2022, Fournier 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

  • 481
    Page views
  • 141
    Downloads
  • 1
    Citations

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

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. Marjorie Fournier
  2. Amélie Rodrigue
  3. Larissa Milano
  4. Jean-Yves Bleuyard
  5. Anthony M Couturier
  6. Jacob Wall
  7. Jessica Ellins
  8. Svenja Hester
  9. Stephen J Smerdon
  10. László Tora
  11. Jean-Yves Masson
  12. Fumiko Esashi
(2022)
KAT2-mediated acetylation switches the mode of PALB2 chromatin association to safeguard genome integrity
eLife 11:e57736.
https://doi.org/10.7554/eLife.57736

Further reading

    1. Chromosomes and Gene Expression
    2. Immunology and Inflammation
    Suhas Sureshchandra, Brianna M Doratt ... Ilhem Messaoudi
    Research Article Updated

    Maternal pre-pregnancy (pregravid) obesity is associated with adverse outcomes for both mother and offspring. Amongst the complications for the offspring is increased susceptibility and severity of neonatal infections necessitating admission to the intensive care unit, notably bacterial sepsis and enterocolitis. Previous studies have reported aberrant responses to LPS and polyclonal stimulation by umbilical cord blood monocytes that were mediated by alterations in the epigenome. In this study, we show that pregravid obesity dysregulates umbilical cord blood monocyte responses to bacterial and viral pathogens. Specifically, interferon-stimulated gene expression and inflammatory responses to respiratory syncytial virus (RSV) and E. coli were significantly dampened, respectively . Although upstream signaling events were comparable, translocation of the key transcription factor NF-κB and chromatin accessibility at pro-inflammatory gene promoters following TLR stimulation was significantly attenuated. Using a rhesus macaque model of western style diet-induced obesity, we further demonstrate that this defect is detected in fetal peripheral monocytes and tissue-resident macrophages during gestation. Collectively, these data indicate that maternal obesity alters metabolic, signaling, and epigenetic profiles of fetal monocytes leading to a state of immune paralysis during late gestation and at birth.

    1. Chromosomes and Gene Expression
    2. Plant Biology
    Myeongjune Jeon, Goowon Jeong ... Ilha Lee
    Research Article

    To synchronize flowering time with spring, many plants undergo vernalization, a floral-promotion process triggered by exposure to long-term winter cold. In Arabidopsis thaliana, this is achieved through cold-mediated epigenetic silencing of the floral repressor, FLOWERING LOCUS C (FLC). COOLAIR, a cold-induced antisense RNA transcribed from the FLC locus, has been proposed to facilitate FLC silencing. Here, we show that C-repeat (CRT)/dehydration-responsive elements (DREs) at the 3′-end of FLC and CRT/DRE-binding factors (CBFs) are required for cold-mediated expression of COOLAIR. CBFs bind to CRT/DREs at the 3′-end of FLC, both in vitro and in vivo, and CBF levels increase gradually during vernalization. Cold-induced COOLAIR expression is severely impaired in cbfs mutants in which all CBF genes are knocked-out. Conversely, CBF-overexpressing plants show increased COOLAIR levels even at warm temperatures. We show that COOLAIR is induced by CBFs during early stages of vernalization but COOLAIR levels decrease in later phases as FLC chromatin transitions to an inactive state to which CBFs can no longer bind. We also demonstrate that cbfs and FLCΔCOOLAIR mutants exhibit a normal vernalization response despite their inability to activate COOLAIR expression during cold, revealing that COOLAIR is not required for the vernalization process.