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© The Rockefeller University Press, 0021-9525/2000//405 $5.00
The Journal of Cell Biology, Volume 148, Number 3, , 2000 405-416

A Role for Centrin 3 in Centrosome Reproduction

^a Institut Curie, Section Recherche, UMR 144 du CNRS, 75248 Paris Cedex 05, France
^b Département de Biologie Moléculaire et Structurale, Laboratoire du Cytosquelette, INSERM U366, Centre d'Études Nucléaires, 38041 Grenoble Cedex, France
Institut Curie-Recherche, UMR 144 du CNRS, 26, rue d'Ulm, 75248 Paris Cedex 05, France.01 42 34 64 2101 42 34 64 20

mbornens{at}curie.fr

Centrosome reproduction by duplication is essential for the bipolarity of cell division, but the molecular basis of this process is still unknown. Mutations in Saccharomyces cerevisiae CDC31 gene prevent the duplication of the spindle pole body (SPB). The product of this gene belongs to the calmodulin super-family and is concentrated at the half bridge of the SPB. We present a functional analysis of HsCEN3, a human centrin gene closely related to the CDC31 gene. Tran- sient overexpression of wild-type or mutant forms of HsCen3p in human cells demonstrates that centriole localization depends on a functional fourth EF-hand, but does not produce mitotic phenotype. However, injection of recombinant HsCen3p or of RNA encoding HsCen3p in one blastomere of two-cell stage Xenopus laevis embryos resulted in undercleavage and inhibition of centrosome duplication. Furthermore, HsCEN3 does not complement mutations or deletion of CDC31 in S. cerevisiae, but specifically blocks SPB duplication, indicating that the human protein acts as a dominant negative mutant of CDC31. Several lines of evidence indicate that HsCen3p acts by titrating Cdc31p-binding protein(s).

Our results demonstrate that, in spite of the large differences in centrosome structure among widely divergent species, the centrosome pathway of reproduction is conserved.

Key Words: centrosome • duplication • Ca²⁺-binding protein • yeast • Xenopus laevis

© 2000 The Rockefeller University Press

Abbreviations used in this paper: CEN, centromeric; HsCEN1, Homo sapiens centrin1 (CETN1 in the human genome database); HsCEN2, Homo sapiens centrin2 (CETN2 in the human genome database); HsCEN3, Homo sapiens centrin3 (CETN3 in the human genome database); SPB, spindle pole body.

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