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Title: Functional characterisation of FAM40A and FAM40B as novel regulators of the cytoskeleton
Author: Suryavanshi, Narendra Bhupal
Awarding Body: King's College London
Current Institution: King's College London (University of London)
Date of Award: 2013
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This thesis describes the functional characterisation of the two human FAM40 proteins, FAM40A and FAM40B, in cancer cells and endothelial cells. FAM40A and FAM40B were initially studied in PC3 cells (a prostate cancer cell line) to determine their effects on cell morphology, adhesion and migration. PC3 cells show distinct morphological phenotypes when depleted of FAM40A and FAM40B using RNAi (RNA interference). Knockdown of FAM40A causes cells to have a smaller spread area, but cells depleted of FAM40B are elongated. FAM40-depleted PC3 cells display an adhesion defect, and FAM40B depletion results in a cell spreading defect. Additionally, FAM40B knockdown in PC3 cells leads to a reduction in cell speed. FAM40A and FAM40B were identified as interacting partners for the CCM3 (cerebral cavernous malformation 3) protein in a previous biochemical study using mass spectrometry. CCM3 is mutated in a subset of patients with cerebral cavernous malformations (CCM). Patients display defects in the vascular endothelium of the central nervous system. The FAM40 proteins were found to form homo- and hetero- dimers, and interact with CCM3 and the serine/threonine kinases MST4 and STK25. Moreover, both FAM40 proteins and CCM3 can be phosphorylated in vitro by MST4. Knockdown of FAM40A, FAM40B and CCM3 in endothelial cells by RNAi causes an increase in stress fibres. FAM40B depletion also reduces endothelial barrier function. Furthermore, depletion of FAM40A, FAM40B and CCM3 results in defects in an in vitro angiogenesis assay. These results demonstrate the importance of the FAM40 proteins for endothelial cell physiology, and suggest they act as part of the CCM3-containing protein complex. Taken together, the FAM40 proteins have important roles in cancer cell motility and adhesion, and are regulators of endothelial cell function. In addition to being implicated in CCM, these proteins could regulate two major steps in cancer progression - cancer cell migration, and neovascularisation.
Supervisor: Ridley, Anne ; Jones, Gareth E. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available