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Concentration of intraflagellar transport proteins at the ciliary base is required for proper train injection

Jamin Jung, View ORCID ProfileJulien Santi-Rocca, View ORCID ProfileCécile Fort, View ORCID ProfileJean-Yves Tinevez, Cataldo Schietroma, View ORCID ProfilePhilippe Bastin
doi: https://doi.org/10.1101/2021.08.02.454739
Jamin Jung
1Trypanosome Cell Biology Unit, Institut Pasteur, INSERM U1201, Paris, France
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Julien Santi-Rocca
1Trypanosome Cell Biology Unit, Institut Pasteur, INSERM U1201, Paris, France
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Cécile Fort
1Trypanosome Cell Biology Unit, Institut Pasteur, INSERM U1201, Paris, France
2Sorbonne Université, Ecole Doctorale Complexité du Vivant, Paris, France
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Jean-Yves Tinevez
3UtechS Photonic Bioimaging (Imagopole), Institut Pasteur, Paris, France
4Image Analysis Hub, C2RT, Institut Pasteur, Paris, France
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Cataldo Schietroma
5Abbelight, , Cachan, France
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Philippe Bastin
1Trypanosome Cell Biology Unit, Institut Pasteur, INSERM U1201, Paris, France
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  • ORCID record for Philippe Bastin
  • For correspondence: pbastin@pasteur.fr
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Abstract

Construction of cilia and flagella relies on Intraflagellar Transport (IFT). Although IFT proteins can be found in multiple locations in the cell, transport has only been reported along the axoneme. Here, we reveal that IFT concentration at the base of the flagellum of Trypanosoma brucei is required for proper assembly of IFT trains. Using live cell imaging at high resolution and direct optical nanoscopy with axially localized detection (DONALD) of fixed trypanosomes, we demonstrate that IFT proteins are localised around the 9 doublet microtubules of the proximal portion of the transition zone, just on top of the transition fibres. Super-resolution microscopy and photobleaching studies reveal that knockdown of the RP2 transition fibre protein results in reduced IFT protein concentration and turnover at the base of the flagellum. This in turn is accompanied by a 4- to 8-fold drop in the frequency of IFT train injection. We propose that the flagellum base provides a unique environment to assemble IFT trains.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted August 02, 2021.
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Concentration of intraflagellar transport proteins at the ciliary base is required for proper train injection
Jamin Jung, Julien Santi-Rocca, Cécile Fort, Jean-Yves Tinevez, Cataldo Schietroma, Philippe Bastin
bioRxiv 2021.08.02.454739; doi: https://doi.org/10.1101/2021.08.02.454739
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Concentration of intraflagellar transport proteins at the ciliary base is required for proper train injection
Jamin Jung, Julien Santi-Rocca, Cécile Fort, Jean-Yves Tinevez, Cataldo Schietroma, Philippe Bastin
bioRxiv 2021.08.02.454739; doi: https://doi.org/10.1101/2021.08.02.454739

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