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Stem cell biology

Base excision repair proteins couple activation-induced cytidine deaminase and endonuclease G during replication stress-induced MLL destabilization

Leukemia volume 32, pages 159–167 (2018)Cite this article

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Abstract

The breakpoint cluster region of the MLL gene (MLLbcr) is frequently rearranged in therapy-related and infant acute leukaemia, but the destabilizing mechanism is poorly understood. We recently proposed that DNA replication stress results in MLLbcr cleavage via endonuclease G (EndoG) and represents the common denominator of genotoxic therapy-induced MLL destabilization. Here we performed a siRNA screen for new factors involved in replication stress-induced MLL rearrangements employing an enhanced green fluorescent protein-based reporter system. We identified 10 factors acting in line with EndoG in MLLbcr breakage or further downstream in the repair of the MLLbcr breaks, including activation-induced cytidine deaminase (AID), previously proposed to initiate MLLbcr rearrangements in an RNA transcription-dependent mechanism. Further analysis connected AID and EndoG in MLLbcr destabilization via base excision repair (BER) components. We show that replication stress-induced recruitment of EndoG to the MLLbcr and cleavage are AID/BER dependent. Notably, inhibition of the core BER factor Apurinic-apyrimidinic endonuclease 1 protects against MLLbcr cleavage in tumour and human cord blood-derived haematopoietic stem/progenitor cells, harbouring the cells of origin of leukaemia. We propose that off-target binding of AID to the MLLbcr initiates BER-mediated single-stranded DNA cleavage, which causes derailed EndoG activity ultimately resulting in leukaemogenic MLLbcr rearrangements.

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Acknowledgements

We thank Daniela Waldraff, Regina Häfele and Jasmin Petermeier (all from the Department of Obstetrics and Gynaecology, Ulm University, Ulm, Germany) for their help with experimental procedures. This work was financially supported by the German Research Foundation (DFG, WI3099/7-1, WI3099/7-2 and PA3 in Research Training Group 1789 ‘Cellular and Molecular Mechanisms in aging’, CEMMA) and the German Space Agency/German Ministry of Education and Research (DLR/BMBF, project TRANSCAN ERA-NET 01KT1302).

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  1. Department of Obstetrics and Gynaecology, Gynaecological Oncology, Ulm University, Ulm, Germany

    B Gole, E Mian, M Rall & L Wiesmüller

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Gole, B., Mian, E., Rall, M. et al. Base excision repair proteins couple activation-induced cytidine deaminase and endonuclease G during replication stress-induced MLL destabilization. Leukemia 32, 159–167 (2018). https://doi.org/10.1038/leu.2017.191

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