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. 2022 Dec 19;42(8):e111500. doi: 10.15252/embj.2022111500

Figure 3. Suppressor mutations in the Dam1 complex result in elevated spindle assembly checkpoint activity and minichromosome missegregation.

Figure 3

  1. Serial dilutions on YPAD media at the permissive (25°C) or restrictive (35°C) temperatures for the ipl1‐321 mutation. The suppressor mutations from the Dam1c (blue) were the only mutants that rescued viability at the restrictive temperature.
  2. Top: representative images showing Bub3‐mNeonGreen localization to kinetochores (Nuf2‐mRuby3), indicating spindle assembly checkpoint activation in wild‐type yeast. Scale bar is 2 μm. Bottom: quantification of the proportion of prometaphase cells with Bub3‐mNeonGreen foci. Only the mutations in Dam1 complex (blue) show a significant increase in SAC activity. Data are from at least five independent experiments. Means and standard deviations are shown.
  3. Proportion of colonies that grow on plates with restrictive (lacking uracil) versus permissive (YPAD) media after 24 h growth under permissive conditions with a URA3‐containing plasmid. The suppressor mutations in the Dam1c (blue) show a significant reduction of minichromosome segregation fidelity. Data are from three independent experiments. Means and standard deviations are shown.

Data information: (ns) nonsignifcant; (*) P < 0.05; (**) P < 0.01; (***) P < 0.001; (****) P < 0.0001; unpaired t‐test.