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J. Cell Biol., Volume 143, Number 6, December 14, 1998 1575-1589

Centriole Disassembly In Vivo and Its Effect on Centrosome Structure and Function in Vertebrate Cells

Y. Bobinnec,* A. Khodjakov,Dagger L.M. Mir,§ C.L. Rieder,Dagger B. Eddé,parallel and M. Bornens*

* Institut Curie, Section Recherche, UMR144 du Centre National de la Recherche Scientifque (CNRS), 75248 Paris Cedex 05, France; Dagger  Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509; § UMR1772 du CNRS, Institut Gustave Roussy, 94805 Villejuif, France; and parallel  Collège de France, UPR9065 du CNRS, Biochimie Cellulaire, 75005 Paris, France

Glutamylation is the major posttranslational modification of neuronal and axonemal tubulin and is restricted predominantly to centrioles in nonneuronal cells (Bobinnec, Y., M. Moudjou, J.P. Fouquet, E. Desbruyères, B. Eddé, and M. Bornens. 1998. Cell Motil. Cytoskel. 39:223-232). To investigate a possible relationship between the exceptional stability of centriole microtubules and the compartmentalization of glutamylated isoforms, we loaded HeLa cells with the monoclonal antibody GT335, which specifically reacts with polyglutamylated tubulin. The total disappearance of the centriole pair was observed after 12 h, as judged both by immunofluorescence labeling with specific antibodies and electron microscopic observation of cells after complete thick serial sectioning. Strikingly, we also observed a scattering of the pericentriolar material (PCM) within the cytoplasm and a parallel disappearance of the centrosome as a defined organelle. However, centriole disappearance was transient, as centrioles and discrete centrosomes ultimately reappeared in the cell population.

During the acentriolar period, a large proportion of monopolar half-spindles or of bipolar spindles with abnormal distribution of PCM and NuMA were observed. However, as judged by a quasinormal increase in cell number, these cells likely were not blocked in mitosis.

Our results suggest that a posttranslational modification of tubulin is critical for long-term stability of centriolar microtubules. They further demonstrate that in animal cells, centrioles are instrumental in organizing centrosomal components into a structurally stable organelle.

Key words: centrosomecentrioletubulin polyglutamylationmitosismitotic spindle


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