Super-resolution microscopy reveals coupling between mammalian centriole subdistal appendages and distal appendages

Elife. 2020 Apr 3:9:e53580. doi: 10.7554/eLife.53580.

Abstract

Subdistal appendages (sDAPs) are centriolar elements that are observed proximal to the distal appendages (DAPs) in vertebrates. Despite the obvious presence of sDAPs, structural and functional understanding of them remains elusive. Here, by combining super-resolved localization analysis and CRISPR-Cas9 genetic perturbation, we find that although DAPs and sDAPs are primarily responsible for distinct functions in ciliogenesis and microtubule anchoring, respectively, the presence of one element actually affects the positioning of the other. Specifically, we find dual layers of both ODF2 and CEP89, where their localizations are differentially regulated by DAP and sDAP integrity. DAP depletion relaxes longitudinal occupancy of sDAP protein ninein to cover the DAP region, implying a role of DAPs in sDAP positioning. Removing sDAPs alter the distal border of centrosomal γ-tubulins, illustrating a new role of sDAPs. Together, our results provide an architectural framework for sDAPs that sheds light on functional understanding, surprisingly revealing coupling between DAPs and sDAPs.

Keywords: CEP89; Ninein; ODF2; cell biology; developmental biology; distal appendages; human; subdistal appendages; super-resolution microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle
  • Cell Cycle Proteins / chemistry
  • Cells, Cultured
  • Centrioles / ultrastructure*
  • Cytoskeletal Proteins / chemistry
  • Heat-Shock Proteins / chemistry
  • Humans
  • Microscopy, Electron, Transmission / methods*
  • Microtubule-Associated Proteins / chemistry
  • Nuclear Proteins / chemistry

Substances

  • CEP89 protein, human
  • Cell Cycle Proteins
  • Cep170 protein, human
  • Cytoskeletal Proteins
  • Heat-Shock Proteins
  • Microtubule-Associated Proteins
  • NIN protein, human
  • Nuclear Proteins
  • ODF2 protein, human