Distribution of a centrosomal antigen during morphogenesis in the ciliated protozoan Euplotes.
Ciliates assemble basal bodies in great number at many stages of the life-cycle. In order to understand their assembly mechanisms, we screened a library of monoclonal antibodies directed against pericentriolar material. One of these antibodies, CTR210, was used previously to follow steps of this assembly process: in Paraurostyla, new basal bodies appear along a scaffold of linear structures recognized by this antibody. The very unusual behavior of this antigen deserved confirmation in other species. In the present study, we show by immunofluorescence that, in another phylogenetically very distant species, Euplotes, basal bodies are assembled in the same pathway during division. In addition, this antibody recognizes a filamentous ring located at the division furrow and linking many basal body assemblages. By cell fractionation and cytoskeletal extraction, we obtained fractions enriched in basal bodies and associated material. Such fractions still display a high complexity in protein composition. These fractions were used to characterize the main target of the antibody as a doublet of 45 kDa. These results confirm previous results in terms of functionality of the protein recognized by the antibody, but raise new questions in terms of the assignment of the recognized protein to the HSP70 family as hypothesized previously.
Fleury
A
A
Laboratoire de Biologie Cellulaire 4, Université Paris XI, Orsay, France.
Lemullois
M
M
Coffe
G
G
D000818:Animals; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan / Q000378:metabolism*; D002455:Cell Division; D018385:Centrosome / Q000276:immunology* / Q000648:ultrastructure; D016798:Ciliophora / Q000254:growth & development / Q000276:immunology / Q000648:ultrastructure; D016812:Euplotes / Q000254:growth & development* / Q000276:immunology* / Q000648:ultrastructure; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D007120:Immunochemistry; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D008970:Molecular Weight; D015800:Protozoan Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D018840:HSP70 Heat-Shock Proteins; D015800:Protozoan Proteins