SAS-6 engineering reveals interdependence between cartwheel and microtubules in determining centriole architecture

Nat Cell Biol. 2016 Apr;18(4):393-403. doi: 10.1038/ncb3329. Epub 2016 Mar 21.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algal Proteins / chemistry
  • Algal Proteins / genetics
  • Algal Proteins / metabolism*
  • Blotting, Western
  • Cell Cycle Proteins / chemistry
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Cell Line, Tumor
  • Centrioles / chemistry
  • Centrioles / metabolism*
  • Centrioles / ultrastructure
  • Chlamydomonas reinhardtii / genetics
  • Chlamydomonas reinhardtii / metabolism*
  • Crystallography, X-Ray
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Microscopy, Atomic Force
  • Microscopy, Electron
  • Microscopy, Fluorescence
  • Microtubules / chemistry
  • Microtubules / metabolism*
  • Microtubules / ultrastructure
  • Models, Molecular
  • Molecular Conformation
  • Mutation
  • Protein Multimerization
  • Protein Structure, Tertiary
  • RNA Interference

Substances

  • Algal Proteins
  • Cell Cycle Proteins
  • SASS6 protein, human
  • Green Fluorescent Proteins