%0 Journal Article %T Flagellar incorporation of proteins follows at least two different routes in trypanosomes %+ Biologie cellulaire des Trypanosomes - Trypanosome Cell Biology %A Vincensini, Laetitia %A Blisnick, Thierry %A Bertiaux, Eloise %A Hutchinson, Sebastian %A Georgikou, Christina %A Ooi, Cher-Pheng %A Bastin, Philippe %Z L.V. was supported by a Roux post‐doctoral fellowship from the Institut Pasteur. E.B. is supported by fellowships from French National Ministry for Research and Technology (doctoral school CDV515) and from La Fondation pour la Recherche Médicale (FDT20170436836). S. H. is funded by Roux post‐doctoral fellowship from the Institut Pasteur. This work is funded by ANR grants (11‐BSV8‐016 and 14‐CE35‐0009‐01), by a French Government Investissement d'Avenir programme, Laboratoire d'Excellence “Integrative Biology of Emerging Infectious Diseases” (ANR‐10‐LABX‐62‐IBEID) and by La Fondation pour la Recherche Médicale (Equipe FRM DEQ20150734356). %< avec comité de lecture %@ 0248-4900 %J Biology of the Cell %I Wiley %V 110 %N 2 %P 33-47 %8 2018-02 %D 2018 %R 10.1111/boc.201700052 %M 29148062 %K Arginine kinase %K Axoneme %K Cilia and flagella %K Organelle assembly %K Outer dynein arms %Z Life Sciences [q-bio]/Microbiology and Parasitology %Z Life Sciences [q-bio]Journal articles %X 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. %G English %Z We thank Derrick Robinson (Bordeaux) for providing the Mab25 antibody. We thank the UTechS‐PBI (Imagopole) France–BioImaging infrastructure, supported by the French National Research Agency (ANR10‐INSB‐04‐01, Investments for the Future), for advice and access to the microscopy systems and Jean‐Yves Tinevez for training and valuable advice %L pasteur-03112358 %U https://pasteur.hal.science/pasteur-03112358 %~ INSERM %~ PASTEUR %~ ANR %~ FRM %~ TRYPANOSOME-CELL-BIOL