Flagellar incorporation of proteins follows at least two different routes in trypanosomes

Laetitia Vincensini, Thierry Blisnick, Eloise Bertiaux, Sebastian Hutchinson, Christina Georgikou, CHER-PHENG OOI, Philippe Bastin

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Abstract

Background information: Eukaryotic cilia and flagella are sophisticated organelles composed of several hundreds of proteins that need to be incorporated at the right time and the right place during assembly.

Results: Two methods were used to investigate this process in the model protist Trypanosoma brucei: inducible expression of epitope-tagged labelled proteins and fluorescence recovery after photobleaching of fluorescent fusion proteins. This revealed that skeletal components of the radial spokes (RSP3), the central pair (PF16) and the outer dynein arms (DNAI1) are incorporated at the distal end of the growing flagellum. They display low or even no visible turnover in mature flagella, a finding further confirmed by monitoring a heavy chain of the outer dynein arm. In contrast, the membrane-associated protein arginine kinase 3 (AK3) showed rapid turnover in both growing and mature flagella, without particular polarity and independently of intraflagellar transport.

Conclusion: These results demonstrate different modes of incorporation for structural and membrane-associated proteins in flagella.

Significance: The existence of two distinct modes for incorporation of proteins in growing flagella suggests the existence of different targeting machineries. Moreover, the absence of turnover of structural elements supports the view that the length of the mature flagellum in trypanosomes is not modified after assembly.

Keywords: Arginine kinase; Axoneme; Cilia and flagella; Organelle assembly; Outer dynein arms.
Original languageEnglish
Pages (from-to)33 - 47
Number of pages15
JournalBiology of the Cell
Volume110
Issue number2
Early online date16 Nov 2017
DOIs
Publication statusPublished - 1 Feb 2018

Keywords

  • Arginine kinase
  • Axoneme
  • Cilia and flagella
  • Organelle assembly
  • Outer dynein arms

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    Vincensini, L., Blisnick, T., Bertiaux, E., Hutchinson, S., Georgikou, C., OOI, CHER-PHENG., & Bastin, P. (2018). Flagellar incorporation of proteins follows at least two different routes in trypanosomes. Biology of the Cell, 110(2), 33 - 47. https://doi.org/10.1111/boc.201700052