title: Establishment of Centromeric Chromatin by the CENP-A Assembly Factor CAL1 Requires FACT-Mediated Transcription.
abstract: Centromeres are essential chromosomal structures that mediate accurate chromosome segregation during cell division. Centromeres are specified epigenetically by the heritable incorporation of the centromeric histone H3 variant CENP-A. While many of the primary factors that mediate centromeric deposition of CENP-A are known, the chromatin and DNA requirements of this process have remained elusive. Here, we uncover a role for transcription in Drosophila CENP-A deposition. Using an inducible ectopic centromere system that uncouples CENP-A deposition from endogenous centromere function and cell-cycle progression, we demonstrate that CENP-A assembly by its loading factor, CAL1, requires RNAPII-mediated transcription of the underlying DNA. This transcription depends on the CAL1 binding partner FACT, but not on CENP-A incorporation. Our work establishes RNAPII passage as a key step in chaperone-mediated CENP-A chromatin establishment and propagation.
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, USA.
authors.forename: Chin-Chi
authors.initials: CC
authors.lastname: Chen
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, USA.
authors.forename: Sarion
authors.initials: S
authors.lastname: Bowers
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK; Biological Research Centre of the Hungarian Academy of Sciences, Institute of Biochemistry, P.O. Box 521, 6701 Szeged, Hungary.
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Lipinszki
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Palladino
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, USA.
authors.forename: Sarah
authors.initials: S
authors.lastname: Trusiak
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, USA.
authors.forename: Emily
authors.initials: E
authors.lastname: Bettini
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, USA.
authors.forename: Leah
authors.initials: L
authors.lastname: Rosin
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK.
authors.forename: Marcin R
authors.initials: MR
authors.lastname: Przewloka
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge CB2 3EH, UK.
authors.forename: David M
authors.initials: DM
authors.lastname: Glover
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, USA; Institute for Systems Genomics, University of Connecticut, Storrs, CT 06269, USA.
authors.forename: Rachel J
authors.initials: RJ
authors.lastname: O'Neill
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, USA; Institute for Systems Genomics, University of Connecticut, Storrs, CT 06269, USA. Electronic address: barbara.mellone@uconn.edu.
authors.forename: Barbara G
authors.initials: BG
authors.lastname: Mellone
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002503:Centromere / Q000378:metabolism*; D000076247:Centromere Protein A; D002843:Chromatin / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly / Q000502:physiology; D004268:DNA-Binding Proteins / Q000378:metabolism*; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D008938:Mitosis / Q000502:physiology
chemical_list: C558282:Cal1 protein, Drosophila; D002352:Carrier Proteins; D000076247:Centromere Protein A; D002843:Chromatin; C427237:Cid protein, Drosophila; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; C518656:FACT protein, Drosophila; D006657:Histones