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Updated: Aug 20, 2026

Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins
Published on: March 3, 2016
Centriole assembly requires both centriolar and pericentriolar material proteins
Alexander Dammermann1, Thomas Müller-Reichert, Laurence Pelletier
1Ludwig Institute for Cancer Research, Department of Cellular and Molecular Medicine, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA. adammerman@ucsd.edu
Abstract:
Centrioles organize pericentriolar material to form centrosomes and also template the formation of cilia. Despite the importance of centrioles in dividing and differentiated cells, their assembly remains poorly understood at a molecular level. Here, we develop a fluorescence microscopy-based assay for centriole assembly in the 1-cell stage C. elegans embryo. We use this assay to characterize SAS-6, a centriolar protein that we identified based on its requirement for centrosome duplication. We show that SAS-6, a member of a conserved metazoan protein family, is specifically required for new centriole assembly, a result we confirm by electron microscopy. We further use the centriole assembly assay to examine the roles of three pericentriolar material proteins: SPD-5, the kinase aurora-A, and gamma-tubulin. Our results suggest that the pericentriolar material promotes daughter centriole formation by concentrating gamma-tubulin around the parent centriole. Thus, both centriolar and pericentriolar material proteins contribute to centriole assembly.
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