Centrosome linker diversity and its function in centrosome clustering and mitotic spindle formation

The centrosome linker joins the two interphase centrosomes of a cell into one microtubule organizing center. Despite increasing knowledge on linker components, linker diversity in different cell types and their role in cells with supernumerary centrosomes remained unexplored. Here, we identified Ninein as a C-Nap1-anchored centrosome linker component that provides linker function in RPE1 cells while in HCT116 and U2OS cells, Ninein and Rootletin link centrosomes together. In interphase, overamplified centrosomes use the linker for centrosome clustering, where Rootletin gains centrosome linker function in RPE1 cells. Surprisingly, in cells with centrosome overamplification, C-Nap1 loss prolongs metaphase through persistent activation of the spindle assembly checkpoint indicated by BUB1 and MAD1 accumulation at kinetochores. In cells lacking C-Nap1, the reduction of microtubule nucleation at centrosomes and the delay in nuclear envelop rupture in prophase probably cause mitotic defects like multipolar spindle formation and chromosome mis-segregation. These defects are enhanced when the kinesin HSET, which normally clusters multiple centrosomes in mitosis, is partially inhibited indicating a functional interplay between C-Nap1 and centrosome clustering in mitosis.

Automated summary

The centrosome linker plays a crucial role in connecting the interphase centrosomes of a cell into one microtubule organizing center. The diversity of linker components in different cell types and their function in cells with supernumerary centrosomes have not been fully explored. This study identified Ninein as a component of the centrosome linker anchored by C-Nap1, providing linker function in RPE1 cells while Ninein and Rootletin link centrosomes together in HCT116 and U2OS cells. The loss of C-Nap1 in cells with centrosome overamplification leads to prolonged metaphase, mitotic defects, and an enhanced functional interplay between C-Nap1 and centrosome clustering.