Fig. 3: Characterization of Krp1 as a member of the kinetochore complex.

a Testing the essentiality of kinetochore components. Strains were grown overnight in the absence or presence of 0.05 μg/mL doxycycline (DOX) at which point they were spotted in tenfold dilutions (starting from an OD₆₀₀ of 0.5) onto YNB agar alone or supplemented with 50 μg/mL DOX. Plates were photographed after growth for 48 h at 30 °C. b Examining the impact of kinetochore-related genes on C. albicans morphology. Strains were grown overnight as described in (a). Strains were subsequently subcultured to an OD₆₀₀ of 0.1 in YPD in the absence or presence of 0.05 μg/mL DOX as indicated. The wild-type strain in the absence of DOX was treated with 25 mM hydroxyurea (HU) as indicated. Cultures were incubated at 30 °C for 24 h for GRACE strains or 6 h for HU treatment before cells were visualized by microscopy. Experiment was performed in biological duplicate with similar results. c Krp1 localizes to the kinetochore. Strains were subcultured to an OD₆₀₀ of 0.1 in YPD and allowed to grow for 4 h before visualization. Krp1 (green), Dad1 (red), Mtw1 (red), and nuclei (blue) were visualized by fluorescence microscopy. Experiment was performed in biological duplicate with similar results. d AP-MS of affinity-tagged Krp1 identified physically interacting proteins. Cells were grown in YPD at 30 °C, and statistically significant interactions were defined through SAINTexpress analysis compared with an unrelated tagged protein. Nodes are grouped and colored based on GO term annotation. The weight of the edges reflects the fold-change in peptide count of Krp1 relative to an unrelated tagged protein (Eno1) for those interacting partners with a BFDR < 1%. Source data are provided as a Source Data file.