Fin1p over-expression results in Cdc16p disassociation from the SPB.
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(A) Cells expressing the labeled tubulin marker leu1::m-Cherry-atb2 and cdc16-GFP were induced to express fin1 from the medium strength nmt41 promoter . Cells transformed with empty vector served as control. Cells were grown in medium without thiamine for 27h at 25°C. Cells were imaged and the intensity of SPB associated cdc16-GFP signal was analyzed as described in . The panel shows m-Cherry-atb2 leu1::cdc16-GFP(ura4+) cells bearing REP41 or REP41-fin1. The scale bar is 10 μm. (B) The SPB associated signal was determined in interphase cells in each strain. Since REP41-fin1 eventually leads to a mitotic arrest  interphase cells were identified by the presence of an interphasic microtubule array. The box shows 25%-75% range for the population, the line indicates the median. The bars indicate 10% and 90% range for the population, and dots indicate more extreme individual values. The y-axis shows fluorescence intensity on an arbitrary scale. (C) Cells bearing the leu1::cdc16-HA allele were induced to express fin1 (ON) by growing them in defined minimal medium  in the presence (OFF) or absence (ON) of 2mM thiamine. Protein extracts were prepared 27h after induction and analyzed by western blotting using monoclonal antibody 12CA5. The anti-α-tubulin monoclonal antibody TAT-1  was used as a control.