title: Discovery and characterisation of the automethylation properties of PRDM9.
abstract: We have previously characterised the histone lysine methyltransferase properties of PRDM9, a member of the PRDM family of putative transcriptional regulators. PRDM9 displays broad substrate recognition and methylates a range of histone substrates, including octamers, core histone proteins, and peptides. In the present study, we show that PRDM9 performs intramolecular automethylation on multiple lysine residues localised to a lysine-rich region on the post-SET (suppressor of variegation 3-9, enhancer of zeste and trithorax) domain. PRDM9 automethylation is abolished by a single active-site mutation, C321P, also known to disrupt interactions withS-adenosylmethionine. We have taken an initial step towards tool compound generation through rational design of a substrate-mimic, peptidic inhibitor of PRDM9 automethylation. The discovery of automethylation in PRDM9 adds a new dimension to our understanding of PRDM9 enzymology.
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos 03-01, Singapore 138669.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Koh-Stenta
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos 03-01, Singapore 138669.
authors.forename: Anders
authors.initials: A
authors.lastname: Poulsen
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos 03-01, Singapore 138669.
authors.forename: Rong
authors.initials: R
authors.lastname: Li
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos 03-01, Singapore 138669.
authors.forename: John Liang Kuan
authors.initials: JL
authors.lastname: Wee
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos 03-01, Singapore 138669.
authors.forename: Perlyn Zekui
authors.initials: PZ
authors.lastname: Kwek
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos 03-01, Singapore 138669.
authors.forename: Sin Yin
authors.initials: SY
authors.lastname: Chew
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos 03-01, Singapore 138669.
authors.forename: Jianhe
authors.initials: J
authors.lastname: Peng
authors.affiliation: Institute of Molecular and Cell Biology, A*STAR, 61 Biopolis Drive, Singapore 138673.
authors.forename: Liling
authors.initials: L
authors.lastname: Wu
authors.affiliation: Institute of Molecular and Cell Biology, A*STAR, 61 Biopolis Drive, Singapore 138673.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Guccione
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos 03-01, Singapore 138669.
authors.forename: Joma
authors.initials: J
authors.lastname: Joy
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31 Biopolis Way, Nanos 03-01, Singapore 138669 jhill@etc.a-star.edu.sg.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Hill
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D020134:Catalytic Domain; D003001:Cloning, Molecular; D003545:Cysteine / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D011495:Histone-Lysine N-Methyltransferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D008745:Methylation; D051379:Mice; D008958:Models, Molecular; D009154:Mutation; D011392:Proline / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D000072757:Protein Conformation, beta-Strand; D000072417:Protein Domains; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D008024:Ligands; D011994:Recombinant Proteins; D011392:Proline; D011495:Histone-Lysine N-Methyltransferase; C534829:prdm9 protein, mouse; D003545:Cysteine
keywords: automethylation; histone methyltransferase; molecular dynamics; substrate-mimic inhibitor