title: Selective Y centromere inactivation triggers chromosome shattering in micronuclei and repair by non-homologous end joining.
abstract: Chromosome missegregation into a micronucleus can cause complex and localized genomic rearrangements known as chromothripsis, 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.
authors.affiliation: Ludwig Institute for Cancer Research and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Peter
authors.identifier: http://orcid.org/0000-0001-8946-7069
authors.initials: P
authors.lastname: Ly
authors.affiliation: Department of Biology, Massachusetts Institute of Technology and Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA.
authors.forename: Levi S
authors.initials: LS
authors.lastname: Teitz
authors.affiliation: Ludwig Institute for Cancer Research and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Dong H
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Ludwig Institute for Cancer Research and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Ofer
authors.initials: O
authors.lastname: Shoshani
authors.affiliation: Howard Hughes Medical Institute and Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA.
authors.forename: Helen
authors.initials: H
authors.lastname: Skaletsky
authors.affiliation: Ludwig Institute for Cancer Research and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Daniele
authors.identifier: http://orcid.org/0000-0002-8795-6771
authors.initials: D
authors.lastname: Fachinetti
authors.affiliation: Department of Biology, Massachusetts Institute of Technology and Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA.|Howard Hughes Medical Institute and Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA.
authors.forename: David C
authors.initials: DC
authors.lastname: Page
authors.affiliation: Ludwig Institute for Cancer Research and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Don W
authors.identifier: http://orcid.org/0000-0001-5489-6453
authors.initials: DW
authors.lastname: Cleveland
mesh_terms: D001324:Autoantigens / Q000378:metabolism; D045744:Cell Line, Tumor; D002503:Centromere / Q000378:metabolism*; D000076247:Centromere Protein A; D051936:Centromere Protein B / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D020090:Chromosome Segregation; D041322:Chromosomes, Human, Y / Q000378:metabolism*; D000072837:Chromothripsis; D059766:DNA End-Joining Repair; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D048629:Micronuclei, Chromosome-Defective; D008938:Mitosis
chemical_list: D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone