Dynein-2 intermediate chains play crucial but distinct roles in primary cilia formation and function

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Abstract

The dynein-2 microtubule motor is the retrograde motor for intraflagellar transport. Mutations in dynein-2 components cause skeletal ciliopathies, notably Jeune syndrome. Dynein-2 contains a heterodimer of two non-identical intermediate chains, WDR34 and WDR60. Here, we use knockout cell lines to demonstrate that each intermediate chain has a distinct role in cilium function. Using quantitative proteomics, we show that WDR34 KO cells can assemble a dynein-2 motor complex that binds IFT proteins yet fails to extend an axoneme, indicating complex function is stalled. In contrast, WDR60 KO cells do extend axonemes but show reduced assembly of dynein-2 and binding to IFT proteins. Both proteins are required to maintain a functional transition zone and for efficient bidirectional intraflagellar transport. Our results indicate that the subunit asymmetry within the dynein-2 complex is matched with a functional asymmetry between the dynein-2 intermediate chains. Furthermore, this work reveals that loss of function of dynein-2 leads to defects in transition zone architecture, as well as intraflagellar transport.

Data availability

Proteomics data are included as an Excel file which is readily readable and accessible for the majority of researchers. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE [1] partner repository with the dataset identifier PXD010398.

The following data sets were generated

1. Stephens DJ et al
(2018) Mass spectrometry proteomics data
Accession number: PXD010398.

https://www.ebi.ac.uk/pride

Article and author information

Author details

Laura Vuolo

School of Biochemistry, University of Bristol, Bristol, United Kingdom

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-9801-9206
Nicola L Stevenson

School of Biochemistry, University of Bristol, Bristol, United Kingdom

Competing interests
The authors declare that no competing interests exist.
Kate J Heesom

Proteomics Facility, Faculty of Biomedical Sciences, University of Bristol, Bristol, United Kingdom

Competing interests
The authors declare that no competing interests exist.
David John Stephens

School of Biochemistry, University of Bristol, Bristol, United Kingdom

For correspondence
david.stephens@bristol.ac.uk

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-5297-3240

Funding

Medical Research Council (MR/P000177/1)

Nicola L Stevenson
David John Stephens

Biotechnology and Biological Sciences Research Council (BB/N000420/1)

Laura Vuolo
David John Stephens

Medical Research Council (MR/K018019/1)

Nicola L Stevenson
David John Stephens

Biotechnology and Biological Sciences Research Council (BB/L014181/1)

David John Stephens

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.