DNA polymerase theta suppresses mitotic crossing over

English version

Autoři: Juan Carvajal-Garcia ^aff001; K. Nicole Crown ^aff002; Dale A. Ramsden ^aff001; Jeff Sekelsky ^aff001
Působiště autorů: Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, North Carolina, United States of America ^aff001; Department of Biology, Case Western Reserve University, Cleveland, Ohio, United States of America ^aff002; Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina, United States of America ^aff003; Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina, United States of America ^aff004; Integrative Program in Biological and Genome Sciences, University of North Carolina, Chapel Hill, North Carolina, United States of America ^aff005
Vyšlo v časopise: DNA polymerase theta suppresses mitotic crossing over. PLoS Genet 17(3): e1009267. doi:10.1371/journal.pgen.1009267
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pgen.1009267

Souhrn

Polymerase theta-mediated end joining (TMEJ) is a chromosome break repair pathway that is able to rescue the lethality associated with the loss of proteins involved in early steps in homologous recombination (e.g., BRCA1/2). This is due to the ability of polymerase theta (Pol θ) to use resected, 3’ single stranded DNA tails to repair chromosome breaks. These resected DNA tails are also the starting substrate for homologous recombination. However, it remains unknown if TMEJ can compensate for the loss of proteins involved in more downstream steps during homologous recombination. Here we show that the Holliday junction resolvases SLX4 and GEN1 are required for viability in the absence of Pol θ in Drosophila melanogaster, and lack of all three proteins results in high levels of apoptosis. Flies deficient in Pol θ and SLX4 are extremely sensitive to DNA damaging agents, and mammalian cells require either Pol θ or SLX4 to survive. Our results suggest that TMEJ and Holliday junction formation/resolution share a common DNA substrate, likely a homologous recombination intermediate, that when left unrepaired leads to cell death. One major consequence of Holliday junction resolution by SLX4 and GEN1 is cancer-causing loss of heterozygosity due to mitotic crossing over. We measured mitotic crossovers in flies after a Cas9-induced chromosome break, and observed that this mutagenic form of repair is increased in the absence of Pol θ. This demonstrates that TMEJ can function upstream of the Holiday junction resolvases to protect cells from loss of heterozygosity. Our work argues that Pol θ can thus compensate for the loss of the Holliday junction resolvases by using homologous recombination intermediates, suppressing mitotic crossing over and preserving the genomic stability of cells.

Klíčová slova:

Apoptosis – DNA damage – DNA repair – Drosophila melanogaster – Genetic causes of cancer – Guide RNA – Heterozygosity – Homozygosity

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