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Title: The role of the centriolar satellite protein PCM-1 in centrosome function
Author: Dammermann, Alexander
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2003
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In most animal cells, centrosomes are responsible for the nucleation and organisation of the mitotic spindle microtubules and the interphase microtubule network. My work has concentrated on the role of one protein, PCM-1, that localises to small cytoplasmic granules known as ‘centriolar satellites’. These satellites have been observed moving between the centrosome and the surrounding cytoplasm. Although their presence has been correlated with the processes of centrosome duplication and ciliogenesis, their precise significance is unknown. The aim of my thesis was to determine the role of PCM-1 and centriolar satellites in centrosome function. Several approaches were taken to inhibit PCM-1 activity: microinjection of antibodies into cultured cells, overexpression of a PCM-1 deletion mutant, and specific depletion of PCM-1 using RNAi. I found that inhibition or depletion of PCM-1 consistently led to mislocalisation of the centrosomal proteins pericentrin, centrin and ninein. Similar effects were observed upon disruption of the dyein/dynactin microtubule motor complex and prolonged exposure of cells to the microtubule-destabilising drug nocodazole. Furthermore, PCM-1 inhibition resulted in a loss of radial microtubule organisation. However, gamma-tubulin in localisation and microtubule nucleation were unaffected. These data are consistent with a model of PCM-1 as a shuttle protein required for the recruitment of centrosomal proteins involved in microtubule anchoring. Centriolar satellites may thus represent protein complexes in transit to and from the centrosome. In addition, cells with impaired PCM-1 function frequently showed defects in cytokinetics and a failure to enter S phase. How PCM-1 is involved in these processes is unclear. I have laid the groundwork for studies to address these and other outstanding questions by creating a construct to express full-length PCM-1 tagged with green fluorescent protein allowing PCM-1 dynamics to be followed in vivo and by setting up a yeast two-hybrid screen to isolate interactors of PCM-1.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available