Skip to main content
Article thumbnail
Location of Repository

Molecular Dissection of the Centrosome Overduplication Pathway in S-Phase Arrested Cells

By Suzanna Prosser


The formation of a bipolar mitotic spindle is crucial for the even segregation of genetic material into two daughter cells during cell division. Each pole of the spindle is organised by a centrosome, therefore it is paramount that two, and only two, centrosomes are present in the cell at the time that it enters mitosis. The presence of too many centrosomes can lead to the formation of multipolar spindles and the uneven segregation of chromosomes.\ud Indeed, cancer cells frequently display supernumerary centrosomes which may contribute to chromosome missegregation and aneuploidy. Normally, following the completion of mitosis, each cell contains a single centrosome. This then duplicates in a semiconservative, templated manner that is tightly linked to the cell cycle. In some instances supernumerary centrosomes arise through uncoupling of this event from the cell cycle such that centrosomes overduplicate within a single cell cycle. Experimentally, centrosome overduplication can be induced in certain cell types by treatment with drugs, such as hydroxyurea, that inhibit DNA synthesis and thereby provoke an S-phase arrest. This assay has been exploited in CHO, U2OS and p53-/- MEF cells to decipher a molecular pathway for centrosome overduplication using a range of pharmacological inhibitors. Distinct granules containing the protein centrin were identified by fluorescence microscopy as early intermediates in this process and shown to form within the nucleus in a Cdk-dependent manner. These foci are then trafficked from the nucleus to the cytoplasm dependent upon the nuclear export machinery. Here, they recruit modified tubulin, PCM-1 and pericentrin, and resemble centriolar satellites. Microtubules and dynein are required to focus these satellites around the centrosome and the formation of centrioles as recognised by electron microscopy. Finally, Hsp90 is required for the recruitment of γ-tubulin to the newlyformed centrioles to construct functional microtubule organising centres. Significantly, intermediate steps in this pathway show similarities to events associated with the de novo centriole formation pathway and the centriolar and acentriolar pathways of ciliogenesis.\ud Together, this work substantially increases our understanding of how supernumerary centrosomes are generated in cells and identifies key events that may be targeted to prevent centrosome overduplication in cancer cells

Publisher: University of Leicester
Year: 2008
OAI identifier:

Suggested articles


  1. (2002). A mammalian in vitro centriole duplication system: evidence for involvement of CDK2/cyclin E and nucleophosmin/B23 in centrosome duplication. doi
  2. (1997). A yeast heat shock transcription factor (Hsf1) mutant is defective in both Hsc82/Hsp82 synthesis and spindle pole body duplication.
  3. (1999). Absence of Brca2 causes genome instability by chromosome breakage and loss associated with centrosome amplification. doi
  4. (2000). Benzylidene lactam compound, KNK437, a novel inhibitor of acquisition of thermotolerance and heat shock protein induction in human colon carcinoma cells.
  5. (1996). Both cyclin A and cyclin E have S-phase promoting (SPF) activity in Xenopus egg extracts.
  6. (2004). Cell cycle progression after cleavage failure: mammalian somatic cells do not possess a "tetraploidy checkpoint". doi
  7. (2007). Cell cycle progression and de novo centriole assembly after centrosomal removal in untransformed human cells. doi
  8. (1982). Centrioles in the cell cycle. doi
  9. (2005). Centrobin: a novel daughter centriole-associated protein that is required for centriole duplication. doi
  10. (2008). Centrosome duplication proceeds during mimosine-induced G1 cell cycle arrest. doi
  11. (2004). Centrosome maturation and duplication in C. elegans require the coiled-coil protein SPD-2. doi
  12. (2003). Centrosome number is controlled by a centrosome-intrinsic block to reduplication. doi
  13. (2000). Construction of centrosomes and spindle poles by molecular motor-driven assembly of protein particles. doi
  14. (2006). Controlling centrosome number: licenses and blocks. doi
  15. (2006). CP110 cooperates with two calcium-binding proteins to regulate cytokinesis and genome stability. doi
  16. (2004). Cyclin-dependent kinase inhibitor indirubin-3'-oxime selectively inhibits human papillomavirus type 16 E7-induced numerical centrosome anomalies. doi
  17. (1999). Cytoplasmic dynein and dynactin in cell division and intracellular transport. doi
  18. (2001). Cytoplasmic dynein intermediate chain phosphorylation regulates binding to dynactin. doi
  19. (2000). Cytoplasmic dynein-mediated assembly of pericentrin and gamma tubulin onto centrosomes. doi
  20. (1991). De novo formation of centrioles in parthenogenetically activated, diploidized rabbit embryos. doi
  21. (2001). Direct regulation of the centrosome duplication cycle by the p53-p21Waf1/Cip1 pathway. doi
  22. (1999). Dynamic association of proteasomal machinery with the centrosome. doi
  23. (1996). Expression of primary cilia in mammalian cells. doi
  24. (2002). Functional role of epsilon-tubulin in the assembly of the centriolar microtubule scaffold. doi
  25. (2000). GFP-centrin as a marker for centriole dynamics in living cells. doi
  26. (1993). Heat shock alters centrosome organization leading to mitotic dysfunction and cell death. doi
  27. (2002). Human Mps1 kinase is required for the spindle assembly checkpoint but not for centrosome duplication. doi
  28. (2003). Human papillomavirus type 16 E7 oncoprotein can induce abnormal centrosome duplication through a mechanism independent of inactivation of retinoblastoma protein family members. doi
  29. (2001). Human papillomavirus type 16 E7 oncoprotein-induced abnormal centrosome synthesis is an early event in the evolving malignant phenotype. doi
  30. (1995). Identification of a complex between centrin and heat shock proteins in CSF-arrested Xenopus oocytes and dissociation of the complex following oocyte activation. doi
  31. (2004). Induction of centrosome amplification and chromosome instability in human bladder cancer cells by p53 mutation and cyclin E overexpression. doi
  32. (1994). Inhibition of cyclin-dependent kinases by purine analogues. doi
  33. (1994). Inhibition of heat shock protein HSP90-pp60v-src heteroprotein complex formation by benzoquinone ansamycins: essential role for stress proteins in oncogenic transformation. doi
  34. (2002). Loss of p53 and centrosome hyperamplification. doi
  35. (2005). Mammalian cells lack checkpoints for tetraploidy, aberrant centrosome number, and cytokinesis failure. doi
  36. (2006). Mechanism limiting centrosome duplication to once per cell cycle. doi
  37. (2007). Microtubules: an overview. doi
  38. (1999). Mitotic regulators govern progress through steps in the centrosome duplication cycle. doi
  39. (2007). Molecular characterization of centriole assembly in ciliated epithelial cells. doi
  40. (1996). Motors involved in spindle assembly and chromosome segregation. doi
  41. (1991). MPS1 and MPS2: novel yeast genes defining distinct steps of spindle pole body duplication. doi
  42. (1999). Nucleolar protein B23 has molecular chaperone activities. doi
  43. (2003). Part of Ran is associated with AKAP450 at the centrosome: involvement in
  44. (2004). Polo-like kinase-2 is required for centriole duplication in mammalian cells. doi
  45. (1995). Preclinical pharmacologic evaluation of geldanamycin as an antitumor agent. doi
  46. (2007). Preventing the degradation of mps1 at centrosomes is sufficient to cause centrosome reduplication in human cells. doi
  47. (1974). Protein synthesis in salivary glands of Drosophila melanogaster: relation to chromosome puffs. doi
  48. (1982). Purification of the major mammalian heat shock proteins.
  49. (2007). Regulated HsSAS-6 levels ensure formation of a single procentriole per centriole during the centrosome duplication cycle. doi
  50. (2001). Roles for 147 embryonic lethal genes on C.elegans chromosome I identified by RNA interference and video microscopy. doi
  51. (2006). Rootletin interacts with C-Nap1 and may function as a physical linker between the pair of centrioles/basal bodies in cells. doi
  52. (2002). Rootletin, a novel coiled-coil protein, is a structural component of the ciliary rootlet. doi
  53. (2001). Specific phosphorylation of nucleophosmin on Thr(199) by cyclin-dependent kinase 2-cyclin E and its role in centrosome duplication. doi
  54. (2008). sSgo1, a major splice variant of Sgo1, functions in centriole cohesion where it is regulated by Plk1. doi
  55. (2005). Temporal and spatial control of nucleophosmin by the Ran-Crm1 complex in centrosome duplication. doi
  56. (2005). The ciliary rootlet maintains long-term stability of sensory cilia. doi
  57. (2000). The human papillomavirus type 16 E6 and E7 oncoproteins cooperate to induce mitotic defects and genomic instability by uncoupling centrosome duplication from the cell division cycle. doi
  58. (2008). The mammalian SPD-2 ortholog Cep192 regulates centrosome biogenesis. doi
  59. (2001). The mitotic spindle: a self-made machine. doi
  60. (2000). The SCF ubiquitin ligase protein slimb regulates centrosome duplication in Drosophila. doi
  61. (2007). The zebrafish maternal-effect gene cellular atoll encodes the centriolar component sas-6 and defects in its paternal function promote whole genome duplication. doi
  62. (1985). Translocation of nucleolar phosphoprotein B23 (37 kDa/pI 5.1) induced by selective inhibitors of ribosome synthesis. doi

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.