TY - JOUR
T1 - Dynamic de novo heterochromatin assembly and disassembly at replication forks ensures fork stability
AU - Gaggioli, Vincent
AU - Lo, Calvin S.Y.
AU - Reverón-Gómez, Nazaret
AU - Jasencakova, Zuzana
AU - Domenech, Heura
AU - Nguyen, Hong
AU - Sidoli, Simone
AU - Tvardovskiy, Andrey
AU - Uruci, Sidrit
AU - Slotman, Johan A.
AU - Chai, Yi
AU - Gonçalves, João G.S.C.Souto
AU - Manolika, Eleni Maria
AU - Jensen, Ole N.
AU - Wheeler, David
AU - Sridharan, Sriram
AU - Chakrabarty, Sanjiban
AU - Demmers, Jeroen
AU - Kanaar, Roland
AU - Groth, Anja
AU - Taneja, Nitika
N1 - Funding Information:
We thank T. Sixma, K. Luger and X. Zhang for stimulating discussions; H. van Attikum and M. Luijsterberg for kindly providing PAGFP-H2A construct and for sharing technical information; P. Krawczyk for sharing technical information on UNC0642; A. Vindigni for sharing PRIMPOL-overexpression plasmid; and J. Vilstrup Johansen for providing bioinformatics support for analysing ChIP–seq data. Funding was provided by Dutch Cancer Society (NWO) Women in STEM Incentive Grant ENW/01054017/12412, Vidi funding (project no. 114122) and ERC funding (grant no. 101078750/#114168) support to N.T. This study was supported by the Oncode Institute, which is partly financed by the Dutch Cancer Society (R.K.). We thank the Josephine Nefkens Cancer Program for infrastructure support. Danish National Research Foundation to the Center for Epigenetics (DNRF82 to S.S., A.T. O.N.J. and A.G.). Proteomics and mass spectrometry research at University of Southern Denmark was supported by VILLUM Center for Bioanalytical Sciences (VILLUM Foundation grant no. 7292 to O.N.J.) and by PRO-MS, Danish National Mass Spectrometry Platform for Functional Proteomics (grant no. 5072-00007B to O.N.J.).
Funding Information:
We thank T. Sixma, K. Luger and X. Zhang for stimulating discussions; H. van Attikum and M. Luijsterberg for kindly providing PAGFP-H2A construct and for sharing technical information; P. Krawczyk for sharing technical information on UNC0642; A. Vindigni for sharing PRIMPOL-overexpression plasmid; and J. Vilstrup Johansen for providing bioinformatics support for analysing ChIP–seq data. Funding was provided by Dutch Cancer Society (NWO) Women in STEM Incentive Grant ENW/01054017/12412, Vidi funding (project no. 114122) and ERC funding (grant no. 101078750/#114168) support to N.T. This study was supported by the Oncode Institute, which is partly financed by the Dutch Cancer Society (R.K.). We thank the Josephine Nefkens Cancer Program for infrastructure support. Danish National Research Foundation to the Center for Epigenetics (DNRF82 to S.S., A.T. O.N.J. and A.G.). Proteomics and mass spectrometry research at University of Southern Denmark was supported by VILLUM Center for Bioanalytical Sciences (VILLUM Foundation grant no. 7292 to O.N.J.) and by PRO-MS, Danish National Mass Spectrometry Platform for Functional Proteomics (grant no. 5072-00007B to O.N.J.).
Publisher Copyright:
© 2023, The Author(s).
PY - 2023/7
Y1 - 2023/7
N2 - Chromatin is dynamically reorganized when DNA replication forks are challenged. However, the process of epigenetic reorganization and its implication for fork stability is poorly understood. Here we discover a checkpoint-regulated cascade of chromatin signalling that activates the histone methyltransferase EHMT2/G9a to catalyse heterochromatin assembly at stressed replication forks. Using biochemical and single molecule chromatin fibre approaches, we show that G9a together with SUV39h1 induces chromatin compaction by accumulating the repressive modifications, H3K9me1/me2/me3, in the vicinity of stressed replication forks. This closed conformation is also favoured by the G9a-dependent exclusion of the H3K9-demethylase JMJD1A/KDM3A, which facilitates heterochromatin disassembly upon fork restart. Untimely heterochromatin disassembly from stressed forks by KDM3A enables PRIMPOL access, triggering single-stranded DNA gap formation and sensitizing cells towards chemotherapeutic drugs. These findings may help in explaining chemotherapy resistance and poor prognosis observed in patients with cancer displaying elevated levels of G9a/H3K9me3.
AB - Chromatin is dynamically reorganized when DNA replication forks are challenged. However, the process of epigenetic reorganization and its implication for fork stability is poorly understood. Here we discover a checkpoint-regulated cascade of chromatin signalling that activates the histone methyltransferase EHMT2/G9a to catalyse heterochromatin assembly at stressed replication forks. Using biochemical and single molecule chromatin fibre approaches, we show that G9a together with SUV39h1 induces chromatin compaction by accumulating the repressive modifications, H3K9me1/me2/me3, in the vicinity of stressed replication forks. This closed conformation is also favoured by the G9a-dependent exclusion of the H3K9-demethylase JMJD1A/KDM3A, which facilitates heterochromatin disassembly upon fork restart. Untimely heterochromatin disassembly from stressed forks by KDM3A enables PRIMPOL access, triggering single-stranded DNA gap formation and sensitizing cells towards chemotherapeutic drugs. These findings may help in explaining chemotherapy resistance and poor prognosis observed in patients with cancer displaying elevated levels of G9a/H3K9me3.
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U2 - 10.1038/s41556-023-01167-z
DO - 10.1038/s41556-023-01167-z
M3 - Article
C2 - 37414849
AN - SCOPUS:85164110736
SN - 1465-7392
VL - 25
SP - 1017
EP - 1032
JO - Nature Cell Biology
JF - Nature Cell Biology
IS - 7
ER -
