Fig. 1: CPK3 phosphorylates the Arabidopsis actin-depolymerizing factor ADF4. | Nature Communications

Fig. 1: CPK3 phosphorylates the Arabidopsis actin-depolymerizing factor ADF4.

From: Arabidopsis calcium-dependent protein kinase 3 regulates actin cytoskeleton organization and immunity

Fig. 1

In vitro kinase assays using recombinant CPK3, CPK6, CPK28, and ADF4. a CPK3 predominantly phosphorylates ADF4 as compared to CPK6 and CPK28. The receptor-like cytoplasmic Ser/Thr kinases b BIK1 and c PBS1 do not phosphorylate ADF4. d CPK3 predominantly phosphorylates ADF4 at amino acid residues Ser105. a–d Experiments were repeated three times. e Mean ± SD of densitometry scans from three independent biological replicates of the kinase assays shown in d. Data is represented as a percentage of WT ADF4 phosphorylation. Asterisks denote standard deviation of the mean compared to WT ADF4 (one-way ANOVA, *p-value < 0.1; **p-value < 0.01). f Two-dimensional gel electrophoresis analysis of HA-tagged ADF4 expressed in WT Col-0, cpk3-2, the kinase inactive-expressing line cpk3-2/CPK3K107M and CIP-treated WT Col-0 protoplasts. Arrow indicates the direction of isoelectric focusing (IEF). Experiments were repeated two times.

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