title: Heterochromatin boundaries are hotspots for de novo kinetochore formation.
abstract: The centromere-specific histone H3 variant CENH3 (also known as CENP-A) is considered to be an epigenetic mark for establishment and propagation of centromere identity. Pulse induction of CENH3 (Drosophila CID) in Schneider S2 cells leads to its incorporation into non-centromeric regions and generates CID islands that resist clearing from chromosome arms for multiple cell generations. We demonstrate that CID islands represent functional ectopic kinetochores, which are non-randomly distributed on the chromosome and show a preferential localization near telomeres and pericentric heterochromatin in transcriptionally silent, intergenic chromatin domains. Although overexpression of heterochromatin protein 1 (HP1) or increasing histone acetylation interferes with CID island formation on a global scale, induction of a locally defined region of synthetic heterochromatin by targeting HP1-LacI fusions to stably integrated Lac operator arrays produces a proximal hotspot for CID deposition. These data indicate that the characteristics of regions bordering heterochromatin promote de novo kinetochore assembly and thereby contribute to centromere identity.
authors.affiliation: Max Planck Institute of Immunobiology and Epigenetics, Stübeweg 51, 79108 Freiburg, Germany.
authors.forename: Agata M
authors.initials: AM
authors.lastname: Olszak
authors.forename: Dominic
authors.initials: D
authors.lastname: van Essen
authors.forename: António J
authors.initials: AJ
authors.lastname: Pereira
authors.forename: Sarah
authors.initials: S
authors.lastname: Diehl
authors.forename: Thomas
authors.initials: T
authors.lastname: Manke
authors.forename: Helder
authors.initials: H
authors.lastname: Maiato
authors.forename: Simona
authors.initials: S
authors.lastname: Saccani
authors.forename: Patrick
authors.initials: P
authors.lastname: Heun
mesh_terms: D000107:Acetylation; D000818:Animals; D002460:Cell Line; D000076247:Centromere Protein A; D042002:Chromatin Assembly and Disassembly; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D059006:Chromosomes, Insect / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004330:Drosophila / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D006570:Heterochromatin / Q000378:metabolism*; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D018386:Kinetochores / Q000378:metabolism*; D007763:Lac Operon; D056968:Lac Repressors / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016615:Telomere / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D015854:Up-Regulation
chemical_list: D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; C427237:Cid protein, Drosophila; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D006570:Heterochromatin; D006657:Histones; D056968:Lac Repressors; D011993:Recombinant Fusion Proteins; D000090266:Chromobox Protein Homolog 5