title: Variant PRC1 complex-dependent H2A ubiquitylation drives PRC2 recruitment and polycomb domain formation.
abstract: Chromatin modifying activities inherent to polycomb repressive complexes PRC1 and PRC2 play an essential role in gene regulation, cellular differentiation, and development. However, the mechanisms by which these complexes recognize their target sites and function together to form repressive chromatin domains remain poorly understood. Recruitment of PRC1 to target sites has been proposed to occur through a hierarchical process, dependent on prior nucleation of PRC2 and placement of H3K27me3. Here, using a de novo targeting assay in mouse embryonic stem cells we unexpectedly discover that PRC1-dependent H2AK119ub1 leads to recruitment of PRC2 and H3K27me3 to effectively initiate a polycomb domain. This activity is restricted to variant PRC1 complexes, and genetic ablation experiments reveal that targeting of the variant PCGF1/PRC1 complex by KDM2B to CpG islands is required for normal polycomb domain formation and mouse development. These observations provide a surprising PRC1-dependent logic for PRC2 occupancy at target sites in vivo.
authors.affiliation: Laboratory of Chromatin Biology and Transcription, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK.
authors.forename: Neil P
authors.initials: NP
authors.lastname: Blackledge
authors.affiliation: Laboratory of Chromatin Biology and Transcription, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK.
authors.forename: Anca M
authors.initials: AM
authors.lastname: Farcas
authors.affiliation: Laboratory of Developmental Genetics, RIKEN Center for Integrative Medical Sciences, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kondo
authors.affiliation: Laboratory of Chromatin Biology and Transcription, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK.
authors.forename: Hamish W
authors.initials: HW
authors.lastname: King
authors.affiliation: Ubiquitin Proteolysis Group, Central Proteomics Facility, Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, OX3 7BN, UK.
authors.forename: Joanna F
authors.initials: JF
authors.lastname: McGouran
authors.affiliation: Laboratory of Chromatin Biology and Transcription, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK.
authors.forename: Lars L P
authors.initials: LLP
authors.lastname: Hanssen
authors.affiliation: Laboratory of Developmental Genetics, RIKEN Center for Integrative Medical Sciences, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Ito
authors.affiliation: Laboratory of Developmental Epigenetics, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK.
authors.forename: Sarah
authors.initials: S
authors.lastname: Cooper
authors.affiliation: Laboratory of Developmental Genetics, RIKEN Center for Integrative Medical Sciences, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Kaori
authors.initials: K
authors.lastname: Kondo
authors.affiliation: Laboratory of Developmental Genetics, RIKEN Center for Integrative Medical Sciences, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Koseki
authors.affiliation: Laboratory of Developmental Genetics, RIKEN Center for Integrative Medical Sciences, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Ishikura
authors.affiliation: Laboratory of Chromatin Biology and Transcription, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK.
authors.forename: Hannah K
authors.initials: HK
authors.lastname: Long
authors.affiliation: Laboratory of Chromatin Biology and Transcription, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK.
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Sheahan
authors.affiliation: Laboratory of Developmental Epigenetics, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK.
authors.forename: Neil
authors.initials: N
authors.lastname: Brockdorff
authors.affiliation: Ubiquitin Proteolysis Group, Central Proteomics Facility, Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, OX3 7BN, UK.
authors.forename: Benedikt M
authors.initials: BM
authors.lastname: Kessler
authors.affiliation: Laboratory of Developmental Genetics, RIKEN Center for Integrative Medical Sciences, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Koseki
authors.affiliation: Laboratory of Chromatin Biology and Transcription, Department of Biochemistry, University of Oxford, Oxford, OX1 3QU, UK. Electronic address: rob.klose@bioch.ox.ac.uk.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Klose
mesh_terms: D000818:Animals; D001846:Bone Development; D018899:CpG Islands; D053595:Embryonic Stem Cells / Q000378:metabolism*; D044783:F-Box Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005804:Genes, Lethal; D055106:Genome-Wide Association Study; D006657:Histones / Q000378:metabolism*; D056484:Jumonji Domain-Containing Histone Demethylases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D063150:Polycomb Repressive Complex 1 / Q000378:metabolism*; D063151:Polycomb Repressive Complex 2 / Q000378:metabolism*; D017434:Protein Structure, Tertiary
chemical_list: D044783:F-Box Proteins; D006657:Histones; D056484:Jumonji Domain-Containing Histone Demethylases; C524747:Kdm2b protein, mouse; D063151:Polycomb Repressive Complex 2; D063150:Polycomb Repressive Complex 1