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Alternative end-joining is suppressed by the canonical NHEJ component Xrcc4–ligase IV during chromosomal translocation formation

Nature Structural & Molecular Biology volume 17, pages 410–416 (2010)Cite this article

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Abstract

Chromosomal translocations in hematologic and mesenchymal tumors form overwhelmingly by nonhomologous end-joining (NHEJ). Canonical NHEJ, essential for the repair of radiation-induced and some programmed double-strand breaks (DSBs), requires the Xrcc4–ligase IV complex. For other DSBs, the requirement for Xrcc4–ligase IV is less stringent, suggesting the existence of alternative end-joining (alt-NHEJ) pathways. To understand the contributions of the canonical NHEJ and alt-NHEJ pathways, we examined translocation formation in cells deficient in Xrcc4–ligase IV. We found that Xrcc4–ligase IV is not required for but rather suppresses translocations. Translocation breakpoint junctions have similar characteristics in wild-type cells and cells deficient in Xrcc4–ligase IV, including an unchanged bias toward microhomology, unlike what is observed for intrachromosomal DSB repair. Complex insertions in some junctions show that joining can be iterative, encompassing successive processing steps before joining. Our results imply that alt-NHEJ is the primary mediator of translocation formation in mammalian cells.

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Figure 1: Chromosomal translocations are suppressed by Xrcc4–ligase IV.
Figure 2: Translocation breakpoint junctions have similar characteristics in wild-type, Xrcc4−/− mutant, Xrcc4-complemented Xrcc4−/− and Ku70−/− cells.
Figure 3: Insertions at breakpoint junctions show complex characteristics.

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Acknowledgements

We thank M. Leversha and J. McGuire at the Memorial Sloan-Kettering Cancer Center Molecular Cytogenetics Core Facility for performing the FISH analysis, F. Alt and J. Chaudhuri for materials and E. Brunet, D. Weinstock, Y. Akamatsu and other members of Jasin laboratory for helpful discussions. This work was supported by US National Institutes of Health grant R01 NIHGM54668 and a Dorothy Rodbell Cohen Cancer Research Program Grant (M.J.).

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  1. Developmental Biology Program, Memorial Sloan-Kettering Cancer Center and Weill Cornell Graduate School of Medical Sciences, New York, New York, USA

    Deniz Simsek & Maria Jasin

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  1. Deniz Simsek
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D.S. performed the experiments; D.S. and M.J. designed the research and wrote the paper.

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Correspondence to Maria Jasin.

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Simsek, D., Jasin, M. Alternative end-joining is suppressed by the canonical NHEJ component Xrcc4–ligase IV during chromosomal translocation formation. Nat Struct Mol Biol 17, 410–416 (2010). https://doi.org/10.1038/nsmb.1773

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