title: Chromatin-modifying complex component Nurf55/p55 associates with histones H3 and H4 and polycomb repressive complex 2 subunit Su(z)12 through partially overlapping binding sites.
abstract: Drosophila Nurf55 is a component of different chromatin-modifying complexes, including the PRC2 (Polycomb repressive complex 2). Based on the 1.75-Å crystal structure of Nurf55 bound to histone H4 helix 1, we analyzed interactions of Nurf55 (Nurf55 or p55 in fly and RbAp48/46 in human) with the N-terminal tail of histone H3, the first helix of histone H4, and an N-terminal fragment of the PRC2 subunit Su(z)12 using isothermal calorimetry and pulldown experiments. Site-directed mutagenesis identified the binding site of histone H3 at the top of the Nurf55 WD40 propeller. Unmodified or K9me3- or K27me3-containing H3 peptides were bound with similar affinities, whereas the affinity for K4me3-containing H3 peptides was reduced. Helix 1 of histone H4 and Su(z)12 bound to the edge of the β-propeller using overlapping binding sites. Our results show similarities in the recognition of histone H4 and Su(z)12 and identify Nurf55 as a versatile interactor that simultaneously contacts multiple partners.
authors.affiliation: European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
authors.forename: Agnieszka J
authors.initials: AJ
authors.lastname: Nowak
authors.forename: Claudio
authors.initials: C
authors.lastname: Alfieri
authors.forename: Christian U
authors.initials: CU
authors.lastname: Stirnimann
authors.forename: Vladimir
authors.initials: V
authors.lastname: Rybin
authors.forename: Florence
authors.initials: F
authors.lastname: Baudin
authors.forename: Nga
authors.initials: N
authors.lastname: Ly-Hartig
authors.forename: Doris
authors.initials: D
authors.lastname: Lindner
authors.forename: Christoph W
authors.initials: CW
authors.lastname: Müller
mesh_terms: D000818:Animals; D001665:Binding Sites; D018360:Crystallography, X-Ray; D029721:Drosophila Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster; D011495:Histone-Lysine N-Methyltransferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D063151:Polycomb Repressive Complex 2; D063146:Polycomb-Group Proteins; D017433:Protein Structure, Secondary; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D056505:Retinoblastoma-Binding Protein 4 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: C110251:Caf1-55 protein, Drosophila; D029721:Drosophila Proteins; D006657:Histones; D063146:Polycomb-Group Proteins; D012097:Repressor Proteins; D056505:Retinoblastoma-Binding Protein 4; C543390:Su(z)12 protein, Drosophila; D011495:Histone-Lysine N-Methyltransferase; D063151:Polycomb Repressive Complex 2