Cdc5-Dependent Asymmetric Localization of Bfa1 Fine-Tunes Timely Mitotic Exit
Figure 9
Localization and SPOC activity of Bfa14D in cells with misaligned spindle.
(A) The localization of Bfa14D in cells with misaligned spindles. Δdyn1BFA1-GFP (YSK2103) and Δdyn1mCherry-TUB1BFA14D-GFP (YSK2875) cells were synchronized with α-factor and released at 16°C for 24 h. The localization of Bfa14D in Δdyn1mCherry-TUB1Δbfa1 mutant cells was observed by fluorescence microcopy. Bar, 5 µm. (B,C) The ability of Bfa14D mutants to prevent mitotic exit in Δdyn1Δbfa1 and Δbim1Δbfa1 cells. (B) Δdyn1mCherry-TUB1Δbfa1 (YSK2893), Δdyn1mCherry-TUB1BFA1-GFP (YSK2894) and Δdyn1mCherry-TUB1BFA14D-GFP (YSK2875) cells were synchronized with α-factor at 30°C and released at 16°C for 24 h. Cells with indicated phenotypes were quantified (n = 600) after staining with DAPI. (C) Δbim1Δbfa1 (YSK1867), Δbim1BFA1 (YSK2093), and Δbim1BFA14D (YSK2898) were serially diluted on YPAD and incubated at either 25°C or 37°C.