Reconstitution of Nucleosome Demethylation and Catalytic Properties of a Jumonji Histone Demethylase

SummaryJumonji 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.

Graphical AbstractFigure optionsDownload high-quality image (283 K)Download as PowerPoint slideHighlights
► Catalytic domain of JMJD2A (cJMJD2A) removes methyl marks in a distributive manner
► Homogeneously methylated nucleosomes were used as substrates
► Quantitative assay for nucleosome demethylation has been developed