Fig. 6: SUMOylation recruits Cdc48 via its co-factor Ufd1 to assist in rDNA release.

Co-immunoprecipitation of Cdc48 with ^GFPLrs4 (a) or Nur^16HA (b), or of Nur^16HA with ^GFPLrs4 (c) in cells overexpressing WT or mutant Cdc48. Dpm1 served as loading control. d Y2H analysis of conjugation-deficient SUMO (SUMO^AA) or the mutant unable to recognize SIMs (SUMO^FAIA-AA) with Ufd1 and ufd1∆SIM. e Y2H analysis of Csm1 and Lrs4 with Cdc48, Npl4, Ufd1, and ufd1∆SIM. f Y2H analysis of Nur1 C with Ufd1 and ufd1∆SIM. g Rates of rDNA recombination in WT, ufd1∆SIM, lrs4∆ or ufd1∆SIM lrs4∆ cells (left; n = 3), in WT and ufd1∆SIM strains expressing SUMO at endogenous levels or overexpressed from vectors with the TEF1 promoter (SUMO-OE) (right; n = 3 for WT and SUMO-OE; n = 4 for ufd1∆SIM and ufd1∆SIM SUMO-OE). h Rates of rDNA marker loss in nur1∆ and nur1∆ ufd1∆SIM cells transformed with plasmids bearing NUR1 or its phosphomimetic version (nur1Pmim) with the endogenous promoter (left; n = 3), or bearing NUR1 or its phosphomutant version (nur1Pmut) with the GAL1 promoter (right; n = 3). For nur1Pmut, cells were grown in galactose-containing media for 4 days. d–f Fusions with Gal4-activating domain (AD) or Gal4-DNA-binding domain (BD) are indicated. Cells spotted on -LWHA selective medium were grown for 4 days (f). The rate of marker loss is calculated as in Fig. 3; data are mean ± SEM of n independent biological replicates, shown in log₂ scale relative to WT (g) or nur1∆+NUR1 (h). Statistical analysis was performed using ANOVA, and letters denote significant differences with a Tukey’s post hoc test at P ≤ 0.05. Source data are provided as a Source Data file.