title: Evidence that ubiquitylated H2B corrals hDot1L on the nucleosomal surface to induce H3K79 methylation.
abstract: Ubiquitylation of histone H2B at lysine 120 (H2B-Ub), a post-translational modification first discovered in 1980, plays a critical role in diverse nuclear processes including the regulation of transcription and DNA damage repair. Herein, we use a suite of protein chemistry methods to explore how H2B-Ub stimulates hDot1L-mediated methylation of histone H3 on lysine 79 (H3K79me). By using semisynthetic 'designer' chromatin containing H2B-Ub bearing a site-specifically installed photocrosslinker, here we report an interaction between a functional hotspot on ubiquitin and the N-terminus of histone H2A. Our biochemical studies indicate that this interaction is required for stimulation of hDot1L activity and leads to a repositioning of hDot1L on the nucleosomal surface, which likely places the active site of the enzyme proximal to H3K79. Collectively, our data converge on a possible mechanism for hDot1L stimulation in which H2B-Ub physically 'corrals' the enzyme into a productive binding orientation.
authors.affiliation: Department of Chemistry, Princeton University, Princeton, 08544 New Jersey, USA.
authors.forename: Linjiao
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Laboratory of Synthetic Protein Chemistry, The Rockefeller University, New York, 10065 New York, USA.
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Holt
authors.affiliation: Department of Chemistry, Princeton University, Princeton, 08544 New Jersey, USA.
authors.forename: Nami
authors.initials: N
authors.lastname: Ohashi
authors.affiliation: Department of Chemistry, Princeton University, Princeton, 08544 New Jersey, USA.
authors.forename: Aishan
authors.initials: A
authors.lastname: Zhao
authors.affiliation: Department of Chemistry, Princeton University, Princeton, 08544 New Jersey, USA.
authors.forename: Manuel M
authors.initials: MM
authors.lastname: Müller
authors.affiliation: Department of Chemistry, Princeton University, Princeton, 08544 New Jersey, USA.|Laboratory of Synthetic Protein Chemistry, The Rockefeller University, New York, 10065 New York, USA.
authors.forename: Boyuan
authors.initials: B
authors.lastname: Wang
authors.affiliation: Department of Chemistry, Princeton University, Princeton, 08544 New Jersey, USA.
authors.forename: Tom W
authors.initials: TW
authors.lastname: Muir
mesh_terms: D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D011495:Histone-Lysine N-Methyltransferase; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008745:Methylation; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D009707:Nucleosomes; D011485:Protein Binding; D011487:Protein Conformation; D057149:Ubiquitinated Proteins
chemical_list: D006657:Histones; D009707:Nucleosomes; D057149:Ubiquitinated Proteins; C473477:DOT1L protein, human; D008780:Methyltransferases; D011495:Histone-Lysine N-Methyltransferase
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