title: Centriole assembly requires both centriolar and pericentriolar material proteins.
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.
authors.affiliation: 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
authors.forename: Alexander
authors.initials: A
authors.lastname: Dammermann
authors.forename: Thomas
authors.initials: T
authors.lastname: Müller-Reichert
authors.forename: Laurence
authors.initials: L
authors.lastname: Pelletier
authors.forename: Bianca
authors.initials: B
authors.lastname: Habermann
authors.forename: Arshad
authors.initials: A
authors.lastname: Desai
authors.forename: Karen
authors.initials: K
authors.lastname: Oegema
mesh_terms: 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
chemical_list: 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