biostudies-literatureHinshaw SMUniversity of California San DiegoHoward Hughes Medical InstituteUniversity of CaliforniaNational Institutes of HealthHelen Hay Whitney FoundationNIH HHSNIGMS NIH HHS688-696.e6https://www.ebi.ac.uk/biostudies/studies/S-EPMC9992315biostudies-literatureUnknown33(4)Kinetochores control eukaryotic chromosome segregation by connecting chromosomal centromeres to spindle microtubules. Duplication of centromeric DNA necessitates kinetochore disassembly and subsequent reassembly on nascent sisters. To search for a regulatory mechanism that controls the earliest steps of this process, we studied Mif2/CENP-C, an essential basal component of the kinetochore. We found that phosphorylation of a central region of Mif2 (Mif2-PEST) enhances inner kinetochore assembly. Eliminating Mif2-PEST phosphorylation sites progressively impairs cellular fitness. The most severe Mif2-PEST mutations are lethal in cells lacking otherwise non-essential inner kinetochore factors. These data show that multi-site phosphorylation of Mif2/CENP-C controls inner kinetochore assembly.Current biology : CBMulti-site phosphorylation of yeast Mif2/CENP-C promotes inner kinetochore assembly.PMC9992315S10 OD023498OD023498R01 GM116897GM116897Cai JQuan YZhou ALZhou HHinshaw SMfalseMulti-site phosphorylation of yeast Mif2/CENP-C promotes inner kinetochore assembly.Kinetochores control eukaryotic chromosome segregation by connecting chromosomal centromeres to spindle microtubules. Duplication of centromeric DNA necessitates kinetochore disassembly and subsequent reassembly on nascent sisters. To search for a regulatory mechanism that controls the earliest steps of this process, we studied Mif2/CENP-C, an essential basal component of the kinetochore. We found that phosphorylation of a central region of Mif2 (Mif2-PEST) enhances inner kinetochore assembly. Eliminating Mif2-PEST phosphorylation sites progressively impairs cellular fitness. The most severe Mif2-PEST mutations are lethal in cells lacking otherwise non-essential inner kinetochore factors. These data show that multi-site phosphorylation of Mif2/CENP-C controls inner kinetochore assembly.2023-01-01T00:00:00Z2023 Feb2025-04-03T23:49:49.667Z2025-04-03T23:49:49.667ZS-EPMC99923153673632310.1016/j.cub.2023.01.012