Fig. 4.

The Mtw1-Cdc14 SPI is the result of Cdc14 recruitment to kinetochores. (A) Cdc14 is normally localized to the nucleolus and when tagged with GFP can recruit GBP. (B) Mtw1-GBP recruits Cdc14-GFP to the kinetochores, and this results in mitotic defects, such as lagging kinetochores. (C–E) GBP-Cdc14 and the catalytically inactive variants are normally localized to the nucleolus, as shown by colocalization with nucleolar protein Fob1. Inclusion of a GFP-tagged kinetochore protein, such as Mtw1-GFP or Nnf1-GFP, is sufficient to recruit either GBP-Cdc14 (F and G) or phosphatase-dead GBP-Cdc14-C283A (H) to the kinetochore. Cell outlines are indicated with red dashed lines. (Scale bars, 5 µm.)