New ubiquitin-dependent mechanisms regulating the Aurora B-Protein Phosphatase 1 balance in Saccharomyces cerevisiae .

Protein ubiquitylation regulates many cellular processes, including cell division. We report here a novel mutation altering the S. cerevisiae E1 ubiquitin activating enzyme ( uba1-W928R) that suppresses the temperature sensitivity and chromosome loss phenotype of a well-characterized Aurora B mutant ( ip1-2 ). The uba1-W928R mutation increases Histone H3-S10 phosphorylation in the ipl1-2 strain, indicating that uba1-W928R acts by increasing Ipl1 activity and/or reducing the opposing PP1 (Glc7 in S. cerevisiae ) phosphatase activity. Consistent with this hypothesis, Ipl1 protein levels and stability are elevated in the uba1-W928R mutant, likely mediated via the E2 enzymes Ubc4 and Cdc34. In contrast, the uba1-W928R mutation does not affect Glc7 stability, but exhibits synthetic lethality with several glc7 mutations. Moreover, uba1-W928R cells have altered subcellular distribution of Glc7 and form nuclear Glc7 foci. These effects are likely mediated via the E2 enzymes Rad6 and Cdc34. Our new UBA1 allele reveals new roles for ubiquitylation in regulating the Ipl1-Glc7 balance in budding yeast. While ubiquitylation likely regulates Ipl1 protein stability via the canonical pathway of proteasomal degradation, a non-canonical ubiquitin-dependent pathway maintains normal Glc7 localization and activity.