Downloads provided by UsageCountsExpression of MLL-AF4 or AF4-MLL fusions does not impact the efficiency of DNA damage repair
Expression of MLL-AF4 or AF4-MLL fusions does not impact the efficiency of DNA damage repair
The most frequent rearrangement of the human MLL gene fuses MLL to AF4 resulting in high-risk infant B-cell acute lymphoblastic leukemia (B-ALL). MLL fusions are also hallmark oncogenic events in secondary acute myeloid leukemia. They are a direct consequence of mis-repaired DNA double strand breaks (DNA-DSBs) due to defects in the DNA damage response associated with exposure to topoisomerase-II poisons such as etoposide. It has been suggested that MLL fusions render cells susceptible to additional chromosomal damage upon exposure to etoposide. Conversely, the genome-wide mutational landscape in MLL-rearranged infant B-ALL has been reported silent. Thus, whether MLL fusions compromise the recognition and/or repair of DNA damage remains unanswered. Here, the fusion proteins MLL-AF4 (MA4) and AF4-MLL (A4M) were CRISPR/Cas9-genome edited in the AAVS1 locus of HEK293 cells as a model to study MLL fusion-mediated DNA-DSB formation/repair. Repair kinetics of etoposide- and ionizing radiation-induced DSBs was identical in WT, MA4- and A4M-expressing cells, as revealed by flow cytometry, by immunoblot for γH2AX and by comet assay. Accordingly, no differences were observed between WT, MA4- and A4M-expressing cells in the presence of master proteins involved in non-homologous end-joining (NHEJ; i.e.KU86, KU70), alternative-NHEJ (Alt-NHEJ; i.e.LigIIIa, WRN and PARP1), and homologous recombination (HR, i.e.RAD51). Moreover, functional assays revealed identical NHEJ and HR efficiency irrespective of the genotype. Treatment with etoposide consistently induced cell cycle arrest in S/G2/M independent of MA4/A4M expression, revealing a proper activation of the DNA damage checkpoints. Collectively, expression of MA4 or A4M does neither influence DNA signaling nor DNA-DSB repair.
MLL.AF4, DNA End-Joining Repair, Oncogene Proteins, Fusion, AF4.MLL, Gene Expression, Infant, Cell Cycle Checkpoints, Precursor Cell Lymphoblastic Leukemia-Lymphoma, DSB, Histones, HEK293 Cells, Radiation, Ionizing, Humans, DNA Breaks, Double-Stranded, Comet Assay, Infant leukemia, Homologous Recombination, Myeloid-Lymphoid Leukemia Protein, Research Paper, DNA Damage, Etoposide
MLL.AF4, DNA End-Joining Repair, Oncogene Proteins, Fusion, AF4.MLL, Gene Expression, Infant, Cell Cycle Checkpoints, Precursor Cell Lymphoblastic Leukemia-Lymphoma, DSB, Histones, HEK293 Cells, Radiation, Ionizing, Humans, DNA Breaks, Double-Stranded, Comet Assay, Infant leukemia, Homologous Recombination, Myeloid-Lymphoid Leukemia Protein, Research Paper, DNA Damage, Etoposide
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