Supplemental Material

This article contains the following supporting material:

• Figure 1 - Fta2 protein levels are unaltered in spc7-23 and mal2-1 mutant cells. Protein extracts prepared from wild-type, mal2-1 or spc7-23 strains which expressed Fta2-HA endogenously were used for immunoprecipitations using an anti-HA-antibody. The immunoprecipitates were analysed by Western blot analysis using an anti-HA antibody. Strains were grown at 25 °C or shifted to 36 °C for 6 hours. Protein extracts used had very similar protein concentrations. Actin was used as a loading control.

• Figure 2 - Sequence alignment of S. pombe Fta2 and S. cerevisiae Ctf13 proteins. Blue letters denote identical residues, red letters indicate similar amino acids. The amino acid changes found in the Fta2-291 and Fta2-292 mutant proteins are indicated.

• Figure 3 - Fta2-HA co-immunoprecipitates Mal2-GFP. Co-immunoprecipitation of Fta2 and Mal2. Protein extracts from strains expressing Mal2-GFP, Fta2-HA or both were used for immunoprecipitation (IP) with an anti-HA or anti-GFP antibody. The immunoprecipitates were resolved by SDS-Page and probed with an anti-GFP antibody.

• Figure 4 - Mal2-GFP protein levels are unaltered in fta2 mutant cells. Protein extracts prepared from wild-type or fta2-291 strains which expressed Mal2-GFP were used for immunoprecipitations using an anti-GFP-antibody. The immunoprecipitates were analysed by Western blot analysis using an anti-GFP antibody. Strains were grown at 25 °C or shifted to 36 °C for 6 hours.