Centriole Assembly Requires Both Centriolar and Pericentriolar Material Proteins

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 f...

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Bibliographic Details
Published in:Developmental cell 2004-12, Vol.7 (6), p.815-829
Main Authors: Dammermann, Alexander, Müller-Reichert, Thomas, Pelletier, Laurence, Habermann, Bianca, Desai, Arshad, Oegema, Karen
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Aurora Kinases
Blotting, Western
Caenorhabditis elegans
Caenorhabditis elegans Proteins - chemistry
Cell Cycle
Cell Cycle Proteins
Centrioles - metabolism
Centrioles - ultrastructure
Centrosome - ultrastructure
Cloning, Molecular
Computational Biology
Green Fluorescent Proteins - metabolism
HeLa Cells
Humans
Microscopy, Electron, Transmission
Microscopy, Fluorescence - methods
Microscopy, Immunoelectron
Microtubules - metabolism
Mitosis
Models, Biological
Molecular Sequence Data
Prophase
Protein Kinases - metabolism
Protein-Serine-Threonine Kinases
Recombinant Fusion Proteins - metabolism
RNA Interference
Sequence Homology, Amino Acid
Tubulin - metabolism
Xenopus Proteins
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Summary: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 γ-tubulin. Our results suggest that the pericentriolar material promotes daughter centriole formation by concentrating γ-tubulin around the parent centriole. Thus, both centriolar and pericentriolar material proteins contribute to centriole assembly.
ISSN:1534-5807
1878-1551
DOI:10.1016/j.devcel.2004.10.015