FAM35A co-operates with REV7 to coordinate DNA double-strand break repair pathway choice.

Orthwein, A.; Findlay, S.; Heath, J.; Luo, V.; Malina, A.; Li, Z.; Samiei, A.; Cheyou, E. S.; Karam, M.; Bagci, H.; Rahat, D.; Grapton, D.; Lavoie, E. G.; Dove, C.; Khaled, H.; Mann, K.; Klein, K. O.; Tabach, Y.; Cote, J.-F.; Marechal, A.; Morin, T.; Djerir, B.; Park, M. M.; Kuasne, H.

doi:10.1101/365460

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bioRxiv · 10.1101/365460

FAM35A co-operates with REV7 to coordinate DNA double-strand break repair pathway choice.

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Abstract

DNA double-strand breaks (DSBs) can be repaired by two major pathways: non-homologous end-joining (NHEJ) and homologous recombination (HR). DNA repair pathway choice is governed by the opposing activities of 53BP1, in complex with its effectors RIF1 and REV7, and BRCA1. However, it remains unknown how the 53BP1/RIF1/REV7 complex stimulates NHEJ and restricts HR to the S/G2 phases of the cell cycle. Using a mass spectrometry (MS)-based approach, we identify 11 high-confidence REV7 interactors and elucidate the role of a previously undescribed factor, FAM35A/SHDL2, as a novel effector of REV7 in the NHEJ pathway. FAM35A depletion impairs NHEJ-mediated DNA repair and compromises antibody diversification by class switch recombination (CSR) in B-cells. FAM35A accumulates at DSBs in a 53BP1-, RIF1- and REV7-dependent manner and antagonizes HR by limiting DNA end resection. In fact, FAM35A is part of a larger complex composed of REV7 and another previously uncharacterized protein, C20orf196/SHDL1, which promotes NHEJ and limits HR. Together, these results establish FAM35A as a novel effector of REV7 in controlling the decision-making process during DSB repair.

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Orthwein, A., Findlay, S., Heath, J., Luo, V., Malina, A., Li, Z., Samiei, A., Cheyou, E. S., Karam, M., Bagci, H., Rahat, D., Grapton, D., Lavoie, E. G., Dove, C., Khaled, H., Mann, K., Klein, K. O., Tabach, Y., Cote, J.-F., Marechal, A., Morin, T., Djerir, B., Park, M. M., Kuasne, H.. 2018-07-11. FAM35A co-operates with REV7 to coordinate DNA double-strand break repair pathway choice.. https://doi.org/10.1101/365460

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