Plk4-induced centriole biogenesis in human cells

J Kleylein-Sohn, J Westendorf, M Le Clech… - Developmental cell, 2007 - cell.com
J Kleylein-Sohn, J Westendorf, M Le Clech, R Habedanck, YD Stierhof, EA Nigg
Developmental cell, 2007cell.com
We show that overexpression of Polo-like kinase 4 (Plk4) in human cells induces
centrosome amplification through the simultaneous generation of multiple procentrioles
adjoining each parental centriole. This provided an opportunity for dissecting centriole
assembly and characterizing assembly intermediates. Critical components were identified
and ordered into an assembly pathway through siRNA and localized through
immunoelectron microscopy. Plk4, hSas-6, CPAP, Cep135, γ-tubulin, and CP110 were …
Summary
We show that overexpression of Polo-like kinase 4 (Plk4) in human cells induces centrosome amplification through the simultaneous generation of multiple procentrioles adjoining each parental centriole. This provided an opportunity for dissecting centriole assembly and characterizing assembly intermediates. Critical components were identified and ordered into an assembly pathway through siRNA and localized through immunoelectron microscopy. Plk4, hSas-6, CPAP, Cep135, γ-tubulin, and CP110 were required at different stages of procentriole formation and in association with different centriolar structures. Remarkably, hSas-6 associated only transiently with nascent procentrioles, whereas Cep135 and CPAP formed a core structure within the proximal lumen of both parental and nascent centrioles. Finally, CP110 was recruited early and then associated with the growing distal tips, indicating that centrioles elongate through insertion of α-/β-tubulin underneath a CP110 cap. Collectively, these data afford a comprehensive view of the assembly pathway underlying centriole biogenesis in human cells.
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