Abstract
We used intrachromosomal substrates to directly monitor the effect of the cell cycle on the efficiency and the accuracy of nonhomologous end joining (NHEJ) in mammalian cells. We show that both KU and KU-independent (KU-alt) pathways are efficient when maintaining cells in G1/S, in G2/M or during dynamic progression through S phase. In addition, the accuracy of NHEJ is barely altered when the cells are blocked in G1/S or in G2/M. However, progression through S phase increases the frequency of deletions, which is a hallmark of the KU-alt pathway. Moreover, we show that the intermediates that are generated by the KU-dependent end joining of non-fully complementary ends, and which contain mismatches, nicks or gap intermediates, are less accurately processed when the cells progress through S phase. In conclusion, both KU and KU-alt processes are active throughout the cell cycle, but the repair is more error prone during S phase, both by increasing the mutagenic KU-alt pathway and decreasing the accuracy of the repair of the intermediates generated by the KU-dependent pathway.
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Acknowledgements
We thank Dr M Jasin for the I-SceI expression vector plasmid, and C Laulier, Drs P Bertrand, S Marcand, I Plo and D Marsh for critical reading of the manuscript. This work was supported by La Ligue Nationale contre le Cancer ‘Equipe labellisée, La Ligue 2005’, the ‘programme interorganisme ToxNuc’, INCA and ANR.
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Guirouilh-Barbat, J., Huck, S. & Lopez, B. S-phase progression stimulates both the mutagenic KU-independent pathway and mutagenic processing of KU-dependent intermediates, for nonhomologous end joining. Oncogene 27, 1726–1736 (2008). https://doi.org/10.1038/sj.onc.1210807
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DOI: https://doi.org/10.1038/sj.onc.1210807