FIGURE 10.
Effect of sumoylation and GRK5/PLK2 expression on S129A-GFP protein stability. GAL1 promoter shutoff studies. A, Smt3ts yeast cells expressing S129A-GFP with or without GRK5 or PLK2 at permissive temperature (25 °C, +SUMO) and restrictive temperature (30 °C, −SUMO) were induced for 4 h in galactose (αSyn on) and then transferred to glucose-containing medium (αSyn off). Immunoblotting analysis was performed at the indicated time points after promoter shutoff with αSyn antibody and GAPDH antibody as loading control. A representative result is shown from three independent experiments. B, densitometric analysis of the immunodetection of S129A-GFP relative to the GAPDH loading control. Significance of differences was calculated with one-way ANOVA with Bonferroni's multiple comparison test (**, p < 0.01; ##, p < 0.05 versus 0 h (Bonferroni's multiple comparison test).