Three-dimensional structure of the radial spokes reveals heterogeneity and interactions with dyneins in Chlamydomonas flagella

Cynthia F. Barber, Thomas Heuser, Blanca I. Carbajal-González, Vladimir V. Botchkarev, Daniela Nicastro

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Radial spokes (RSs) play an essential role in the regulation of axonemal dynein activity and thus of ciliary and flagellar motility. However, few details are known about the complexes involved. Using cryo-electron tomography and subtomogram averaging, we visualized the three-dimensional structure of the radial spokes in Chlamydomonas flagella in unprecedented detail. Unlike many other species, Chlamydomonas has only two spokes per axonemal repeat, RS1 and RS2. Our data revealed previously uncharacterized features, including two-pronged spoke bases that facilitate docking to the doublet microtubules, and that inner dyneins connect directly to the spokes. Structures of wild type and the headless spoke mutant pf17 were compared to define the morphology and boundaries of the head, including a direct RS1-to-RS2 interaction. Although the overall structures of the spokes are very similar, we also observed some differences, corroborating recent findings about heterogeneity in the docking of RS1 and RS2. In place of a third radial spoke we found an uncharacterized, shorter electron density named "radial spoke 3 stand-in," which structurally bears no resemblance to RS1 and RS2 and is unaltered in the pf17 mutant. These findings demonstrate that radial spokes are heterogeneous in structure and may play functionally distinct roles in axoneme regulation.

Original languageEnglish (US)
Pages (from-to)111-120
Number of pages10
JournalMolecular Biology of the Cell
Volume23
Issue number1
DOIs
StatePublished - Jan 1 2012

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Chlamydomonas
Dyneins
Flagella
Axonemal Dyneins
Electron Microscope Tomography
Axoneme
Microtubules
Head
Electrons

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Three-dimensional structure of the radial spokes reveals heterogeneity and interactions with dyneins in Chlamydomonas flagella. / Barber, Cynthia F.; Heuser, Thomas; Carbajal-González, Blanca I.; Botchkarev, Vladimir V.; Nicastro, Daniela.

In: Molecular Biology of the Cell, Vol. 23, No. 1, 01.01.2012, p. 111-120.

Research output: Contribution to journalArticle

Barber, Cynthia F. ; Heuser, Thomas ; Carbajal-González, Blanca I. ; Botchkarev, Vladimir V. ; Nicastro, Daniela. / Three-dimensional structure of the radial spokes reveals heterogeneity and interactions with dyneins in Chlamydomonas flagella. In: Molecular Biology of the Cell. 2012 ; Vol. 23, No. 1. pp. 111-120.
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