Identification and characterization of components of the Candida albicans inner kinetochore

Despite the significant global health burden posed by Candida albicans, a large proportion of its genome has yet to be characterized. While insights from model yeasts have historically provided valuable functional clues, their utility is approaching its limits given the increasing evidence of divergence across fungal species. The C. albicans inner kinetochore is a poorly characterized cellular structure. In particular, the constitutive centromere-associated network (CCAN) has only four previously known components, likely due to DNA sequence divergence between orthologous complex members from Saccharomyces cerevisiae or Schizosaccharomyces pombe. Here, we leveraged a structure-focused approach to identify seven components of the C. albicans CCAN. Phenotypic characterization of loss-of-function mutants confirmed important roles in fitness and cell cycle progression for various kinetochore subunits. Furthermore, protein interactions identified through affinity purification-mass spectrometry as well as confocal microscopy confirmed the interaction and localization of these predicted kinetochore components with known kinetochore members. Overall, this work significantly enhances our understanding of a key cellular structure in C. albicans and underscores the urgent need for pathogen-specific research to better understand its unique biological mechanisms.