Fig. 3: Tpk2-catalyzed Jhd2 phosphorylation inhibits the H3K4 demethylase activity of Jhd2. | Nature Communications

Fig. 3: Tpk2-catalyzed Jhd2 phosphorylation inhibits the H3K4 demethylase activity of Jhd2.

From: Phosphorylation of Jhd2 by the Ras-cAMP-PKA(Tpk2) pathway regulates histone modifications and autophagy

Fig. 3

a Western blot analysis of H3K4 methylation in WT, Jhd2-S321A, Jhd2-S340A, Jhd2-S321A, S340A, Jhd2-S321D, Jhd2-S340D, and Jhd2-S321D, S340D mutants. b Western blot analysis of H3K4 methylation in WT, Jhd2-S321A, S340A, Jhd2-S321D, S340D, tpk2∆, tpk2∆ Jhd2-S321A, S340A, and tpk2∆ Jhd2-S321D, S340D mutants. c, d Analysis the activity of Jhd2 purified from WT (Jhd2-FLAG), Jhd2-S321A, S340A (Jhd2-S321A, S340A-FLAG), and tpk2∆ (Jhd2-FLAG tpk2∆) on H3K4me3 (1–23) peptide by in vitro histone demethylase assay. e Dephosphorylation of Jhd2 by λ phosphatase enhanced its histone demethylase activity. f The histone demethylase activity of Jhd2 was reduced when cells grown in YP medium containing increasing glucose concentrations. g In vitro phosphorylation of Jhd2 by purified Tpk2 reduced the histone demethylase activity of WT Jhd2 but not Jhd2-S321A, S340A. For a, b, data represent the mean ± SEM of three biological independent experiments. Two-sided t-tests were used for statistical analysis. Source data are provided as a Source data file.

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