Identification of vinculin as a pericentriolar component in mammalian cells.
Previous studies have shown that centrosome position and structure can be influenced by actin filaments, that centrosomes can influence actin organization, and that an actin homologue is associated with centrosomes. Such observations suggest the existence of connections between centrosomes and actin networks. In keeping with such observations, we show that the pericentriolar material, a main component of centrosomes, contains vinculin, a well-known component of cell adhesion plaques and of adherens cell junctions. We find that in various cell types, centrosomes are specifically stained by five different anti-vinculin antibodies. In adherent cell lines, these antibodies also stained adhesion plaques, but in thymocytes, a cell type devoid of adhesive structures, such antibodies stained only centrosomes. Isolated centrosomes also reacted with the anti-vinculin antibodies and immunoelectron microscopy showed apparent localization of vinculin in the pericentriolar material. Immunoblot analysis confirmed the presence of vinculin in purified centrosomal protein preparations. In such protein fractions, anti-vinculin antibodies reacted with a single polypeptide with an apparent molecular weight similar to that of vinculin. Stepwise solubilization of centrosomal structures using urea showed that high urea concentrations were required to solubilize centrosomal vinculin, suggesting tight association of vinculin with the pericentriolar material. The identification of vinculin as a component of centrosomes provides a possible molecular basis for interaction between F-actin and centrosomes.
Chevrier
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Département de Biologie Moléculaire et Structurale, INSERM U366, Grenoble, France.
Paintrand
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Koteliansky
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Block
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MR
Job
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D000818:Animals; D002417:Cattle; D002458:Cell Fractionation; D002460:Cell Line; D002502:Centrioles / Q000737:chemistry* / Q000648:ultrastructure; D018385:Centrosome / Q000737:chemistry* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D016596:Vinculin / Q000032:analysis*
D016596:Vinculin