Figure 5. Swd2.2, Ppn1 and PP1^Dis2 associate as a protein module to the CPF.

A. The interaction between Flag-tagged Swd2.2 and GFP-tagged Pfs2 was analyzed by co-immunoprecipitation in protein extracts prepared from cycling cells in the presence or absence of Ppn1. Whole cell extracts (WCE) and the immuno-precipitated material (GFP IP) were analyzed by western blot. BCDE. The proteins indicated at the top were purified by affinity and their associated partners were identified by MS/MS mass-spectrometry analysis (see Methods). The number of unique peptides recovered for each protein is indicated. F. Scheme summarizing the proteomic data. GH. Cells expressing the indicated epitope-tagged proteins were synchronized in early mitosis (mitotic) or not (cycling), using the cold-sensitive nda3KM311 mutation [33]. G. Flag-tagged Swd2.2 was immuno-precipitated to look at its interaction with the core CPF component Pfs2. H. Flag-tagged PP1^Dis2 was immuno-precipitated to look at its interaction with the DPS component Swd2.2.