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Title: Tetherin in cytolinesis
Author: Blakemore, Katie Beatrice
Awarding Body: University of Bristol
Current Institution: University of Bristol
Date of Award: 2013
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The work presented in this thesis has examined the localisation and potential role of tetherin during cytokinesis. Tetherin is an integral membrane protein with an unusual topology: an N-terminal cytosolic domain that interacts with the actin cytoskeleton; a transmembrane domain; a large extracellular domain containing an extended parallel coiled -coil, at which the protein dimerises; and a C-terminal glycosylphosphatidylinositol anchor that localises the protein to membrane microdomains. Both GFP-tetherin and endogenous tetherin have recently been observed at the cleavage furrow and midbody of dividing cells. Preliminary evidence also suggested that depletion of tetherin causes failure of cell division, leading to the hypothesis that tetherin may play a role in abscission. The localisation and potential role of tetherin was investigated using confocal microscopy and siRNA depletion. Although robust localisation of tetherin to both the midbody and spindle poles was observed at cytokinesis, depletion of tetherin did not lead to any effect on the rate or efficiency of cell division. Protein interactions with tetherin at cytokinesis were also investigated using proteomics, leading to the discovery of a potential interaction between tetherin and the RhoAlocalising protein Anillin in dividing cells. In addition to investigation of the localisation of tetherin during cytokinesis, the tetherininteracting protein RICH2 was also investigated in the context of cell division. RICH2 could not be observed at the midbody, but proteomic analysis revealed interaction between RICH2 and several members of the Myosin-II family of contractile proteins as well as several regulatory proteins. Depletion of RICH2 was also found to affect the lifetime of midbodies after cell division. Laurdan microscopy was also employed to study the midbody, which was found to exhibit a higher level of lipid organisation than the surrounding plasma membrane. However, this level of organisation was not disrupted by depletion of tetherin.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available