Abstract
ABSTRACTCentrioles are duplicated once in every cell cycle, ensuring the bipolarity of the mitotic spindle. How the core components cooperate to achieve high fidelity in centriole duplication remains poorly understood. By live-cell imaging of endogenously tagged proteins in human cells throughout the entire cell cycle, we quantitatively tracked the dynamics of the critical duplication factors: Plk4, STIL and HsSAS6. Centriolar Plk4 peaks and then starts decreasing during the late G1 phase, which coincides with the accumulation of STIL at centrioles. Shortly thereafter, the HsSAS6 level increases steeply at the procentriole assembly site. We also show that both STIL and HsSAS6 are necessary for attenuating Plk4 levels. Furthermore, our mathematical modeling and simulation suggest that the STIL-HsSAS6 complex in the cartwheel has a negative feedback effect on centriolar Plk4. Combined, these findings illustrate how the dynamic behavior of and interactions between critical duplication factors coordinate the centriole-duplication process.This article has an associated First Person interview with the first author of the paper.
Highlights
Centrosomes and their core components, centrioles, are duplicated once during the cell cycle, which allows bipolar spindle assembly and ensures the proper proliferation of most animal cells (Nigg and Holland, 2018; Gönczy and Hatzopoulos, 2019)
Three centriolar proteins, namely Polo-like kinase 4 (Plk4), STIL and HsSAS6, have been identified as the core components that coordinate the onset of centriole duplication (Banterle and Gönczy, 2017; Nigg and Holland, 2018)
Distinct time courses of centriolar accumulation of endogenous Plk4, STIL and HsSAS6 during the cell cycle To track the behavior of endogenous proteins in live cells, we observed HCT116 cell lines by spinning disc confocal microscopy, as previously described (Takao et al, 2019)
Summary
Centrosomes and their core components, centrioles, are duplicated once during the cell cycle, which allows bipolar spindle assembly and ensures the proper proliferation of most animal cells (Nigg and Holland, 2018; Gönczy and Hatzopoulos, 2019). RESULTS AND DISCUSSION Distinct time courses of centriolar accumulation of endogenous Plk4, STIL and HsSAS6 during the cell cycle To track the behavior of endogenous proteins in live cells, we observed HCT116 cell lines by spinning disc confocal microscopy, as previously described (Takao et al, 2019).
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