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Title: Characterisation of the function(s) of a novel gene, CLWD
Author: Chau, You-Ying
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2007
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The aim of my PhD is to investigate the function(s) of CLWD by characterising its location in cells and in tissues, as well as the phenotype of cells where CLWD is knocked down using siRNA. Results from indirect immunofluoresence suggest that CLWD may be a new component in the mitotic spindle. CLWD localises to centrosomes in prometaphase and moves to mitotic spindles in metaphase. Results from immunohistochemistry show that CLWD is expressed in a wide range of tissues with strong expression in the epithelial areas of tissues. In addition, CLWD is located on human chromosome 20q13.3 and this region is known to be amplified in several cancers. Real-Time PCR analysis shows that CLWD is however not overexpressed in the breast and ovarian tumours analysed. Like 10% of the genes present in human genome, CLWD has an upstream open reading frame (uORF) present in its 5’UTR. Using the luciferase reporter system, I showed that the UORF in CLWD has an inhibitory effect on CLWD translation. I also described some preliminary studies to assess the circadian rhythm regulated nature of Clwd in mouse liver and SCN. Results from RNAi studies suggest that CLWD has a vital role in cell survival. Cells where CLWD is knocked down using siRNA show a decrease in cell proliferation rate, and an increase in mitotic index, suggesting CLWD is important for cell growth. CLWD-depleted cells also show a striking phenotype in nuclei where the nuclei are irregular/lobular in shape (nuclear blebbing). Micronuclei, lagging chromosomes, and multinuclear nuclei are also seen in siClwd transfected cells, suggesting CLWD may play an important role in both mitosis and cytokinesis. Together, I have provided evidence suggesting CLWD may have a crucial role in cell growth and mitosis.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available