RPM-1 Uses Both Ubiquitin Ligase and Phosphatase-Based Mechanisms to Regulate DLK-1 during Neuronal Development
Figure 6
PPM-2 regulates DLK-1 by acting on S874.
(A) Quantitation of PLM axon termination defects (hook) caused by transgenic overexpression of DLK-1 and phosphomimetic DLK-1 point mutants. Note that coexpression of PPM-2 rescues defects caused by overexpression of DLK-1, but not defects caused by overexpression of DLK-1 S874E S878E and DLK-1 S874E. Shown are averages of data pooled from 5 or more transgenic lines for the indicated genotypes; young adult worms (16–20 hours post L4) grown at 23°C were analyzed. (B) CoIP from transgenic whole worm lysates showing that FLAG::DLK-1L K162R coprecipitates with GFP::DLK-1S, and binding of DLK-1L to DLK-1S is not altered in ppm-2-/- mutants (upper panel). (C) Quantitation of FLAG::DLK-1L K162R coIP with GFP::DLK-1S for the indicated genotypes normalized to amount of GFP::DLK-1S precipitated. Shown are the average levels of FLAG::DLK-1 K162R coprecipitating with GFP::DLK-1S acquired from 3 independently derived transgenic lines for each genotype. Error bars represent the standard error of the mean, and significance was determined using an unpaired t-test. **p<0.01, ***p<0.001, ns = not significant.
