title: Structural analysis of the core COMPASS family of histone H3K4 methylases from yeast to human.
abstract: Histone H3 lysine 4 (H3K4) methylation is catalyzed by the highly evolutionarily conserved multiprotein complex known as Set1/COMPASS or MLL/COMPASS-like complexes from yeast to human, respectively. Here we have reconstituted fully functional yeast Set1/COMPASS and human MLL/COMPASS-like complex in vitro and have identified the minimum subunit composition required for histone H3K4 methylation. These subunits include the methyltransferase C-terminal SET domain of Set1/MLL, Cps60/Ash2L, Cps50/RbBP5, Cps30/WDR5, and Cps25/Dpy30, which are all common components of the COMPASS family from yeast to human. Three-dimensional (3D) cryo-EM reconstructions of the core yeast complex, combined with immunolabeling and two-dimensional (2D) EM analysis of the individual subcomplexes reveal a Y-shaped architecture with Cps50 and Cps30 localizing on the top two adjacent lobes and Cps60-Cps25 forming the base at the bottom. EM analysis of the human complex reveals a striking similarity to its yeast counterpart, suggesting a common subunit organization. The SET domain of Set1 is located at the juncture of Cps50, Cps30, and the Cps60-Cps25 module, lining the walls of a central channel that may act as the platform for catalysis and regulative processing of various degrees of H3K4 methylation. This structural arrangement suggested that COMPASS family members function as exo-methylases, which we have confirmed by in vitro and in vivo studies.
authors.affiliation: Stowers Institute for Medical Research, Kansas City, MO 64110, USA.
authors.forename: Yoh-hei
authors.initials: YH
authors.lastname: Takahashi
authors.forename: Gerwin H
authors.initials: GH
authors.lastname: Westfield
authors.forename: Austin N
authors.initials: AN
authors.lastname: Oleskie
authors.forename: Raymond C
authors.initials: RC
authors.lastname: Trievel
authors.forename: Ali
authors.initials: A
authors.lastname: Shilatifard
authors.forename: Georgios
authors.initials: G
authors.lastname: Skiniotis
mesh_terms: D000818:Animals; D017124:Conserved Sequence; D020285:Cryoelectron Microscopy / Q000379:methods; D019175:DNA Methylation; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase / Q000737:chemistry* / Q000235:genetics*; D006657:Histones / Q000737:chemistry*; D006801:Humans; D021621:Imaging, Three-Dimensional; D007313:Insecta; D008745:Methylation; D008854:Microscopy, Electron / Q000379:methods; D008958:Models, Molecular; D008968:Molecular Conformation; D051788:Myeloid-Lymphoid Leukemia Protein / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000235:genetics
chemical_list: D006657:Histones; C497422:KMT2A protein, human; D011994:Recombinant Proteins; D051788:Myeloid-Lymphoid Leukemia Protein; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase