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Endonuclease G initiates DNA rearrangements at the MLL breakpoint cluster upon replication stress

Oncogene volume 34, pages 3391–3401 (2015)Cite this article

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Abstract

MLL (myeloid/lymphoid or mixed-lineage leukemia) rearrangements are frequent in therapy-related and childhood acute leukemia, and are associated with poor prognosis. The majority of the rearrangements fall within a 7.3-kb MLL breakpoint cluster region (MLLbcr), particularly in a 0.4-kb hotspot at the intron11-exon12 boundary. The underlying mechanisms are poorly understood, though multiple pathways including early apoptotic signaling, accompanied by high-order DNA fragmentation, have been implicated. We introduced the MLLbcr hotspot in an EGFP-based recombination reporter system and demonstrated enhancement of both spontaneous and genotoxic treatment-induced DNA recombination by the MLLbcr in various human cell types. We identified Endonuclease G (EndoG), an apoptotic nuclease, as an essential factor for MLLbcr-specific DNA recombination after induction of replication stress. We provide evidence for replication stress-induced nuclear accumulation of EndoG, DNA binding by EndoG as well as cleavage of the chromosomal MLLbcr locus in a manner requiring EndoG. We demonstrate additional dependency of MLLbcr breakage on ATM signaling to histone H2B monoubiquitinase RNF20, involved in chromatin relaxation. Altogether our findings provide a novel mechanism underlying MLLbcr destabilization in the cells of origin of leukemogenesis, with replication stress-activated, EndoG-mediated cleavage at the MLLbcr, which may serve resolution of the stalled forks via recombination repair, however, also permits MLL rearrangements.

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Acknowledgements

We thank Daniela Waldraff, Regina Häfele, Rahel Wiehe (all Department of Obstetrics and Gynecology, University of Ulm, Germany) and Dré van der Merwe (Core facility FACS, Medical Faculty, University of Ulm, Germany) for their contributions to this work. This work was supported by the German Research Foundation (WI 3099/7-1, WI 3099/7-2) and by the European/German Space Agency (ESA/DLR) and German Ministry of Economy (BMWi), A0-10-IBER-2 funding 50WB1225.

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  1. B Gole and C Baumann: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Gynecological Oncology, University of Ulm, Ulm, Germany

    B Gole, C Baumann, E Mian, C I Ireno & L Wiesmüller

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  1. B Gole
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Correspondence to B Gole or L Wiesmüller.

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Gole, B., Baumann, C., Mian, E. et al. Endonuclease G initiates DNA rearrangements at the MLL breakpoint cluster upon replication stress. Oncogene 34, 3391–3401 (2015). https://doi.org/10.1038/onc.2014.268

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