Fig. 5: Tpk2-catalyzed Jhd2 phosphorylation enhances Not4-mediated Jhd2 ubiquitination and its turnover by the proteasome pathway. | Nature Communications

Fig. 5: Tpk2-catalyzed Jhd2 phosphorylation enhances Not4-mediated Jhd2 ubiquitination and its turnover by the proteasome pathway.

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

Fig. 5: Tpk2-catalyzed Jhd2 phosphorylation enhances Not4-mediated Jhd2 ubiquitination and its turnover by the proteasome pathway.

a Western blot analysis of Jhd2 in WT, Jhd2-S321A, S340A, and tpk2Δ mutants. b Analysis of Jhd2 stability in WT, Jhd2-S321A, S340A, and tpk2Δ mutants. c, d Analysis of the effect of MG132 or inactivation of Cim3 on Jhd2 protein levels. e Analysis of the ubiquitination of Jhd2 in WT, Jhd2-S321A, S340A, not4Δ, tpk2Δ, not4Δ Jhd2-S321A, S340A, and not4Δ tpk2Δ mutants. f Analysis of Jhd2, Rpd3 and H3K4 methylation in WT, Jhd2-S321A, S340A, tpk2Δ, not4Δ, Jhd2-S321A, S340A not4Δ, and tpk2Δ not4Δ mutants. g Co-IP assay showing Jhd2-S321A, S340A mutation reduced the interaction between Jhd2 and Not4. h Tpk2-catalyzed Jhd2 phosphorylation increased the interaction between Not4 and Jhd2. i Proposed model showing that Tpk2-catalyzed Jhd2 phosphorylation promotes Not4-mediated ubiquitination and subsequent proteasome degradation of Jhd2. For a–d, f, data represent the mean ± SEM of at least 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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