A human centrosomal protein is immunologically related to basal body-associated proteins from lower eucaryotes and is involved in the nucleation of microtubules.
Isolation of centrosomes from human cells has revealed a proteic pattern which is both complex and specific. As the most prominent structural element of centrosomes in animal cells, the centriole which is present as two copies, is a highly conserved structure, we have attempted to identify centrosomal proteins on the basis of immunocross-reaction with proteins identified in basal bodies from lower eucaryotes. We report that two antibodies, one raised against the Ca(+)-binding protein centrin (Salisbury, J. L., A. T. Baron, B. Surek, and M. Melkonian. 1984. J. Cell Biol. 99:962-970) and the other directed against a 230-kD protein isolated from the infraciliary cytoskeletal lattice of the protozoan Polyplastron m., decorate the centrosome of human cultured cells, and identify one of the major centrosomal components revealed as a doublet of 62/64 kD. Moreover the nucleation reaction of microtubules, which can be efficiently produced on isolated centrosomes, is blocked by the antibodies, a result which strongly implicates the 62/64-kD protein in this centrosomal activity. We also show that the 62/64-kD protein remains insoluble in conditions (0.5 M KI or 8 M urea) which are capable of extracting most of the centrosomal proteins. Immunocytochemical localization by EM of isolated centrosomes revealed the association of this 62/64-kD doublet with the intercentriolar link and the pericentriolar lattice. Our results suggest that conservation of structure in the centrosome from divergent organisms could be matched by conservation of proteins and activity, evidence for the maintenance of a specific function, which could involve Ca2+, associated with the microtubule organizing centers.
Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
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Moudjou
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Paintrand
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Vigues
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Bornens
D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000276:immunology / Q000378:metabolism / Q000502:physiology; D002460:Cell Line; D002502:Centrioles / Q000737:chemistry* / Q000502:physiology; D002868:Chromosomal Proteins, Non-Histone; D002923:Cilia / Q000737:chemistry* / Q000502:physiology; D003285:Contractile Proteins / Q000276:immunology / Q000378:metabolism; D003429:Cross Reactions; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000276:immunology* / Q000502:physiology; D008870:Microtubules / Q000502:physiology*; D008970:Molecular Weight; D010449:Peptide Mapping; D010940:Plant Proteins / Q000276:immunology / Q000378:metabolism; D011485:Protein Binding; D015800:Protozoan Proteins / Q000378:metabolism; D012995:Solubility; D014357:Trypsin
D002135:Calcium-Binding Proteins; D002868:Chromosomal Proteins, Non-Histone; D003285:Contractile Proteins; D008869:Microtubule-Associated Proteins; D010940:Plant Proteins; D015800:Protozoan Proteins; C056608:caltractin; D014357:Trypsin; D002118:Calcium