A Role for Centrin 3 in Centrosome Reproduction

• Published in: The Journal of cell biology Vol. 148; no. 3; pp. 405 - 415
• Main Authors: Middendorp, Sandrine, Küntziger, Thomas, Abraham, Yann, Holmes, Simon, Bordes, Nicole, Paintrand, Michel, Paoletti, Anne, Bornens, Michel
• Format: Journal Article
• Language: English
• Published: United States Rockefeller University Press 07.02.2000; The Rockefeller University Press
• Subjects: Amino acids; Animals; Antibodies; Blastomeres; Calcium-Binding Proteins - genetics; Calcium-Binding Proteins - metabolism; Calcium-Binding Proteins - physiology; CDC31 gene; Cell division; Cells; Cellular biology; Centrin 3; Centrioles - metabolism; Centrosome - physiology; Centrosomes; Embryos; Genes; HeLa Cells; HsCEN3 gene; HsCen3 protein; Humans; Mutation; Original; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; RNA; Saccharomyces cerevisiae; Somatic cells; Spindle pole body; Xenopus laevis; Yeast; Yeasts
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.148.3.405

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. Transient 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.