Selective Y centromere inactivation triggers chromosome shattering in micronuclei and repair by non-homologous end joining

Ly et al. establish a method to selectively inactivate the centromere of the Y chromosome to follow chromosome shattering and micronuclei formation through several cell cycles, and suggest re-ligation of chromosome fragments is dependent on non-homologous end joining. Chromosome missegregation into a micronucleus can cause complex and localized genomic rearrangements 1 , 2 known as chromothripsis 3 , but the underlying mechanisms remain unresolved. Here we developed an inducible Y centromere-selective inactivation strategy by exploiting a CENP-A/histone H3 chimaera to directly examine the fate of missegregated chromosomes in otherwise diploid human cells. Using this approach, we identified a temporal cascade of events that are initiated following centromere inactivation involving chromosome missegregation, fragmentation, and re-ligation that span three consecutive cell cycles. Following centromere inactivation, a micronucleus harbouring the Y chromosome is formed in the first cell cycle. Chromosome shattering, producing up to 53 dispersed fragments from a single chromosome, is triggered by premature micronuclear condensation prior to or during mitotic entry of the second cycle. Lastly, canonical non-homologous end joining (NHEJ), but not homology-dependent repair, is shown to facilitate re-ligation of chromosomal fragments in the third cycle. Thus, initial errors in cell division can provoke further genomic instability through fragmentation of micronuclear DNAs coupled to NHEJ-mediated reassembly in the subsequent interphase.