title: A Basic Post-SET Extension of NSDs Is Essential for Nucleosome Binding In Vitro.
abstract: The nuclear receptor SET domain-containing family of proteins (NSD1, NSD2, and NSD3) is known to mono- and dimethylate lysine 36 of histone H3 (H3K36). Overexpression and translocation of NSDs have been widely implicated in a variety of diseases including cancers. Although the substrate specificity of NSDs has been a subject of many valuable studies, the activity of these proteins has never been fully characterized in vitro. In this study, we present full kinetic characterization of NSD1, NSD2, and NSD3 and provide robust in vitro assays suitable for screening these proteins in a 384-well format using nucleosome as a substrate. Through monitoring the changes in substrate specificity of a series of NSD constructs and using molecular modeling, we show that a basic post-SET extension common to all three NSDs (corresponding to residues 1209 to 1226 of NSD2) is essential for proper positioning on nucleosome substrates.
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Abdellah
authors.initials: A
authors.lastname: Allali-Hassani
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Kuznetsova
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Taraneh
authors.initials: T
authors.lastname: Hajian
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Hong
authors.initials: H
authors.lastname: Wu
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Ludmila
authors.initials: L
authors.lastname: Dombrovski
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Yanjun
authors.initials: Y
authors.lastname: Li
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Susanne
authors.initials: S
authors.lastname: Gräslund
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada Ontario Cancer Institute, The Campbell Family Institute for Cancer Research and Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Cheryl H
authors.initials: CH
authors.lastname: Arrowsmith
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada Department of Pharmacology and Toxicology, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Schapira
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada Department of Pharmacology and Toxicology, University of Toronto, Toronto, Ontario, Canada m.vedadi@utoronto.ca.
authors.forename: Masoud
authors.initials: M
authors.lastname: Vedadi
mesh_terms: D002021:Buffers; D003001:Cloning, Molecular; D017353:Gene Deletion; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase / Q000737:chemistry*; D006657:Histones / Q000737:chemistry; D006801:Humans; D020128:Inhibitory Concentration 50; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry*; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D008958:Models, Molecular; D009154:Mutation; D009687:Nuclear Proteins / Q000737:chemistry*; D009707:Nucleosomes / Q000737:chemistry*; D011485:Protein Binding; D012097:Repressor Proteins / Q000737:chemistry*; D013379:Substrate Specificity
chemical_list: D002021:Buffers; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D009707:Nucleosomes; D012097:Repressor Proteins; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; C113272:NSD1 protein, human; C419586:NSD2 protein, human; C432968:NSD3 protein, human; D008239:Lysine
keywords: MMSET; Sotos syndrome; WHSC1; WHSC1L1; cancer; histone methyltransferase