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

Abstract

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. image Centrosomes are during interphase joined by linkers whose diversity remains unexplored. This work identifies Ninein as a centrosome linker protein that functionally overlaps with Rootletin and shows how the loss of the key linker protein C‐Nap1 affects centrosome functions particularly in cells with extra centrosomes. Ninein has a dual role in centrosome cohesion as centrosome linker component and microtubule anchor. Overamplified centrosomes use the linker for centrosome clustering. C‐Nap1 loss reduces microtubule nucleation and delays nuclear envelope rupture, resulting in mitotic defects like multipolar spindle formation and chromosome mis‐segregation. C‐Nap1 loss prolongs metaphase through persistent activation of the spindle assembly checkpoint in cells with supernumerary centrosomes.