title: SAS-6 engineering reveals interdependence between cartwheel and microtubules in determining centriole architecture.
abstract: Centrioles are critical for the formation of centrosomes, cilia and flagella in eukaryotes. They are thought to assemble around a nine-fold symmetric cartwheel structure established by SAS-6 proteins. Here, we have engineered Chlamydomonas reinhardtii SAS-6-based oligomers with symmetries ranging from five- to ten-fold. Expression of a SAS-6 mutant that forms six-fold symmetric cartwheel structures in vitro resulted in cartwheels and centrioles with eight- or nine-fold symmetries in vivo. In combination with Bld10 mutants that weaken cartwheel-microtubule interactions, this SAS-6 mutant produced six- to eight-fold symmetric cartwheels. Concurrently, the microtubule wall maintained eight- and nine-fold symmetries. Expressing SAS-6 with analogous mutations in human cells resulted in nine-fold symmetric centrioles that exhibited impaired length and organization. Together, our data suggest that the self-assembly properties of SAS-6 instruct cartwheel symmetry, and lead us to propose a model in which the cartwheel and the microtubule wall assemble in an interdependent manner to establish the native architecture of centrioles.
authors.affiliation: Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Manuel
authors.initials: M
authors.lastname: Hilbert
authors.affiliation: Department of Biological Sciences, University of Tokyo, Tokyo 113-0033, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Noga
authors.affiliation: Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Frey
authors.affiliation: Swiss Institute for Experimental Cancer Research (ISREC), School of Life Sciences, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland.
authors.forename: Virginie
authors.initials: V
authors.lastname: Hamel
authors.affiliation: Swiss Institute for Experimental Cancer Research (ISREC), School of Life Sciences, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland.
authors.forename: Paul
authors.initials: P
authors.lastname: Guichard
authors.affiliation: Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Sebastian H W
authors.initials: SH
authors.lastname: Kraatz
authors.affiliation: Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule (ETH) Zürich, CH-4058 Basel, Switzerland.
authors.forename: Moritz
authors.initials: M
authors.lastname: Pfreundschuh
authors.affiliation: Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Sarah
authors.initials: S
authors.lastname: Hosner
authors.affiliation: Swiss Institute for Experimental Cancer Research (ISREC), School of Life Sciences, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Flückiger
authors.affiliation: Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Rolf
authors.initials: R
authors.lastname: Jaussi
authors.affiliation: Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Mara M
authors.initials: MM
authors.lastname: Wieser
authors.affiliation: Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Thieltges
authors.affiliation: Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.|Condensed Matter Theory Group, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Xavier
authors.identifier: http://orcid.org/0000-0003-4572-9316
authors.initials: X
authors.lastname: Deupi
authors.affiliation: Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule (ETH) Zürich, CH-4058 Basel, Switzerland.
authors.forename: Daniel J
authors.identifier: http://orcid.org/0000-0003-3075-0665
authors.initials: DJ
authors.lastname: Müller
authors.affiliation: Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Kammerer
authors.affiliation: Swiss Institute for Experimental Cancer Research (ISREC), School of Life Sciences, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland.
authors.forename: Pierre
authors.initials: P
authors.lastname: Gönczy
authors.affiliation: Department of Biological Sciences, University of Tokyo, Tokyo 113-0033, Japan.
authors.forename: Masafumi
authors.identifier: http://orcid.org/0000-0002-2497-4227
authors.initials: M
authors.lastname: Hirono
authors.affiliation: Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
authors.forename: Michel O
authors.initials: MO
authors.lastname: Steinmetz
mesh_terms: D020418:Algal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002502:Centrioles / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism*; D018360:Crystallography, X-Ray; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D018625:Microscopy, Atomic Force; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D008958:Models, Molecular; D008968:Molecular Conformation; D009154:Mutation; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D034622:RNA Interference
chemical_list: D020418:Algal Proteins; D018797:Cell Cycle Proteins; C497202:SASS6 protein, human; D049452:Green Fluorescent Proteins