Figure 8.

Phosphorylation of Pti4 by Pto Affects Its DNA Binding Activity.

Components as indicated in each lane were preincubated at 25°C for 30 min in 10 μL of kinase buffer. For each kinase reaction, 0.25 μg of His-Pti4 and kinase was used. The final concentration of unlabeled ATP was 1 mM. After incubation, the volume in each tube was adjusted to 30 μL, with the mobility shift assay buffer containing the radiolabeled probe, and assayed as described in Methods. The (+) signs indicate that the corresponding protein, or ATP, was present in the reaction. The position of the unbound (Free) probe is shown.