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 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.
Dammermann
Alexander
A
Ludwig 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
Müller-Reichert
Thomas
T
Pelletier
Laurence
L
Habermann
Bianca
B
Desai
Arshad
A
Oegema
Karen
K
D000595:Amino Acid Sequence; D000818:Animals; D064127:Aurora Kinases; D015153:Blotting, Western; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D002502:Centrioles / Q000378:metabolism / Q000648:ultrastructure*; D018385:Centrosome / Q000648:ultrastructure; D003001:Cloning, Molecular; D019295:Computational Biology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence / Q000379:methods; D016253:Microscopy, Immunoelectron; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis; D008954:Models, Biological; D008969:Molecular Sequence Data; D011418:Prophase; D011494:Protein Kinases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014404:Tubulin / Q000378:metabolism; D029867:Xenopus Proteins
D029742:Caenorhabditis elegans Proteins; D018797:Cell Cycle Proteins; D011993:Recombinant Fusion Proteins; D014404:Tubulin; D029867:Xenopus Proteins; D049452:Green Fluorescent Proteins; D011494:Protein Kinases; C112134:AURKA protein, Xenopus; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases