Characterization, cloning and immunolocalization of a coronin homologue in Trichomonas vaginalis. - Université Clermont Auvergne Accéder directement au contenu
Article Dans Une Revue European Journal of Cell Biology Année : 2000

Characterization, cloning and immunolocalization of a coronin homologue in Trichomonas vaginalis.

Résumé

On adhesion to host cells the flagellate Trichomonas vaginalis switches to an amoeboid form rich in actin microfilaments. We have undertaken the identification of actin-associated proteins that regulate actin dynamics. A monoclonal antibody 4C12 raised against a cytoskeletal fraction of T. vaginalis labeled a protein doublet at circa 50 kDa. These two bands were recognized by the antibody against Dictyostelium discoideum coronin. During cell extraction and actin polymerization, T. vaginalis coronin cosedimented with F-actin. By two-dimensional gel electrophoresis, the protein doublet was separated into two sets of isoforms covering two Ip zones around 6 and 7. By screening a T. vaginalis library with 4C12, two clones Cor 1 and Cor 2 were isolated. This gene duplicity is a particularity among unicellular organisms examined. The complete sequence of the gene Cor 1 encodes a 435-residue protein with a calculated molecular mass of 48 kDa and Ip of 5.58. The incomplete sequence Cor 2 was very similar but with a more basic calculated Ip than Cor 1 on the same region. T. vaginalis coronin had 50% similarity with the coronin family, possessing the five WD-repeats and a leucine zipper in its C-terminal part. Double immunofluorescence labeling showed that coronin mainly colocalized with actin at the periphery of the adherent amoeboid cells. However, coronin labeling displayed patches within a reticular array. Immunogold electron microscopy confirmed the coronin labeling in the actin-rich microfilamentous fringe beneath the plasma membrane, with accumulation in phagocytic zones and pseudopodial extensions. In T. vaginalis, one of the first emerging lineage of eukaryotes, coronin seems to play an important role in actin dynamics and may be a downstream target of a signaling mechanism for the cytoskeleton reorganization.

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Protistologie

Dates et versions

hal-00783606 , version 1 (01-02-2013)

Identifiants

Citer

Geneviève Bricheux, Gérard Coffe, Danielle Bayle, Guy Brugerolle. Characterization, cloning and immunolocalization of a coronin homologue in Trichomonas vaginalis.. European Journal of Cell Biology, 2000, 79 (6), pp.413-22. ⟨10.1078/0171-9335-00065⟩. ⟨hal-00783606⟩
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