Figure 8. Mis4^I803M suppresses pph3Δ psm3^NN chromosome segregation defects.

(A) Cell growth assay. The mis4 mutations restore pph3Δ psm3^NN growth at elevated temperature (B) Model structure of Mis4 showing the location of the mutated residues. (C) Sequence alignment of Mis4 with Scc2 proteins from other species. The mutated residues in Mis4 are colored as in (B). Ct Scc2 residues required for Scc1 binding are indicated by open red circles and Hs Scc2 residues mutated in Cornelia de Lange syndrome are indicated by green dots as in Kikuchi et al. (2016). Ct, Chaetomium thermophilum; Sc, Saccharomyces cerevisiae; Sp, Schizosaccharomyces pombe, Hs, Homo sapiens. (D) Mis4^I803M efficiently suppressed pph3Δ psm3^NN chromosome segregation defects. Cells were arrested in G1 by nitrogen starvation at 25°C and released at 37°C. Progression in the cell cycle was monitored by DNA content analysis. Cells from the 5.75 and 6.25 time points were processed for DNA and tubulin staining to score the number of anaphase cells with lagging chromatids. Bar = 5µm.