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

Autor: Vincensini, Laetitia, Blisnick, Thierry, Bertiaux, Eloise, Hutchinson, Sebastian, Georgikou, Christina, Ooi, Cher-Pheng, Bastin, Philippe
Přispěvatelé: Biologie cellulaire des Trypanosomes - Trypanosome Cell Biology, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris], 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)., 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, ANR-11-BSV8-0016,CCOD,Organisation et dynamique des centrioles et cils(2011), ANR-14-CE35-0009,DECODIFT,Molecular bases for the role of IFT172 in ciliogenesis and in ciliopathy(2014), ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Hutchinson, Sebastian, BLANC - Organisation et dynamique des centrioles et cils - - CCOD2011 - ANR-11-BSV8-0016 - BLANC - VALID, Appel à projets générique - Molecular bases for the role of IFT172 in ciliogenesis and in ciliopathy - - DECODIFT2014 - ANR-14-CE35-0009 - Appel à projets générique - VALID, Integrative Biology of Emerging Infectious Diseases - - IBEID2010 - ANR-10-LABX-0062 - LABX - VALID
Jazyk: angličtina
Rok vydání: 2018
Předmět:
MESH: Signal Transduction
Axoneme
Recombinant Fusion Proteins
MESH: Biological Transport
[SDV]Life Sciences [q-bio]
Trypanosoma brucei brucei
Protozoan Proteins
MESH: Flagella
Cilia and flagella
MESH: Recombinant Fusion Proteins
[SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology
MESH: Protozoan Proteins
Staining and Labeling
Outer dynein arms
Membrane Proteins
MESH: Trypanosoma brucei brucei
Biological Transport
Axonemal Dyneins
MESH: Gene Expression Regulation
Arginine kinase
[SDV] Life Sciences [q-bio]
MESH: Arginine Kinase
MESH: Staining and Labeling
[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology
Gene Expression Regulation
Flagella
Organelle assembly
MESH: Axonemal Dyneins
MESH: Membrane Proteins
MESH: Fluorescence Recovery After Photobleaching
Fluorescence Recovery After Photobleaching
Signal Transduction
Zdroj: Biology of the Cell
Biology of the Cell, Wiley, 2018, 110 (2), pp.33-47. ⟨10.1111/boc.201700052⟩
Biology of the Cell, 2018, 110 (2), pp.33-47. ⟨10.1111/boc.201700052⟩
ISSN: 0248-4900
1768-322X
DOI: 10.1111/boc.201700052⟩
Popis: International audience; 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.
Databáze: OpenAIRE
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