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

Leukemia. 2018 Jan;32(1):159-167. doi: 10.1038/leu.2017.191. Epub 2017 Jun 19.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cells, Cultured
  • Cytidine Deaminase / genetics*
  • DNA Breaks, Double-Stranded
  • DNA Repair / genetics*
  • DNA Replication / genetics*
  • Endodeoxyribonucleases / genetics*
  • Hematopoietic Stem Cells / metabolism
  • Histone-Lysine N-Methyltransferase / genetics*
  • Humans
  • K562 Cells
  • Leukemia / genetics
  • Leukemia / metabolism
  • Myeloid-Lymphoid Leukemia Protein / genetics*
  • Stem Cells / metabolism
  • Transcription, Genetic / genetics

Substances

  • KMT2A protein, human
  • Myeloid-Lymphoid Leukemia Protein
  • Histone-Lysine N-Methyltransferase
  • Endodeoxyribonucleases
  • endonuclease G
  • Cytidine Deaminase