Figure 2.

Gfa1 is required for cyclin C relocalization and destruction. (a) Diagram of Gfa1 functions in the cell. (b) Gfa1 is required for cyclin C destruction. Wild type (RSY10), gfa1-1 cnc1Δ (RSY1543), and chs3Δ (RSY1539) midlog phase cultures expressing myc-cyclin C (pLR337) were treated with 0.4 mM H₂O₂ for the indicated times in hours. Cyclin C levels were determined by Western blot analysis of immunoprecipitates. Tub1 levels were used as a loading control. (c) YFP-cyclin C subcellular localization was monitored in a wild type and gfa1-1 cnc1Δ strains harboring pBK37 before and following H₂O₂ treatment. DAPI staining was used to identify the nucleus. (d) Gfa1 functions upstream of the CWI pathway. Cyclin C levels were determined by Western blot analysis of immunoprecipitates in a midlog phase wild type (RSY10) or gfa1-1 cnc1Δ (RSY1543) cultures expressing myc-cyclin C (pLR337) and either a vector control (v), BCK1-20 or RHO1 ^G19V. Tub1 levels were used as a loading control.