title: Reconstitution of nucleosome demethylation and catalytic properties of a Jumonji histone demethylase.
abstract: Jumonji histone demethylases catalyze removal of methyl marks from lysine residues in histone proteins within nucleosomes. Here, we show that the catalytic domain of demethylase JMJD2A (cJMJD2A) utilizes a distributive mechanism to remove the histone H3 lysine 9 trimethyl mark. By developing a method to assess demethylation of homogeneous, site-specifically methylated nucleosomes, we determined that the kinetic parameters for demethylation of nucleosomes by cJMJD2A are comparable to those of peptide substrates. These findings imply that other domains of the demethylase or its protein partners may contribute to nucleosome recognition in vivo and, in this way, may further regulate demethylation activity and processivity. The quantitative assays of nucleosome demethylation developed in our work provide a platform for future work with complex chromatin substrates and full-length demethylases.
authors.affiliation: Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: Carrie
authors.initials: C
authors.lastname: Shiau
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Trnka
authors.forename: Alen
authors.initials: A
authors.lastname: Bozicevic
authors.forename: Idelisse
authors.initials: I
authors.lastname: Ortiz Torres
authors.forename: Bassem
authors.initials: B
authors.lastname: Al-Sady
authors.forename: Alma L
authors.initials: AL
authors.lastname: Burlingame
authors.forename: Geeta J
authors.initials: GJ
authors.lastname: Narlikar
authors.forename: Danica Galonić
authors.initials: DG
authors.lastname: Fujimori
mesh_terms: D055162:Biocatalysis; D020134:Catalytic Domain; D002843:Chromatin / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006801:Humans; D056484:Jumonji Domain-Containing Histone Demethylases / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D009707:Nucleosomes / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D002843:Chromatin; D006657:Histones; D009707:Nucleosomes; D056484:Jumonji Domain-Containing Histone Demethylases