Date of Award

2014

Publication Type

Master Thesis

Degree Name

M.Sc.

Department

Biological Sciences

First Advisor

Andrew Hubberstey

Keywords

actin binding proteins, actin cytoskeleton, cell migration, F/G actin ratio, mutations

Rights

info:eu-repo/semantics/openAccess

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Abstract

WDR1 is a highly conserved regulator of the actin cytoskeleton in eukaryotes. A novel splice variant of WDR1 has been discovered, WDRΔ35, which lacks exons 3-5. To determine if functional differences exist between these two isoforms and whether perturbation of WDR1/WDRΔ35 levels affects the rate of cell migration, they were overexpressed as GFP fusion proteins in HEK293 cells and the rates of cell migration were quantified. Overexpression of WDR1/WDRΔ35 caused a significant decrease in cell migration compared to the GFP control. The ratio of G/F-actin was measured upon WDR1/WDRΔ35 overexpression and it was found to be significantly higher for GFP-WDR/ WDRΔ35 transfected cells compared to GFP alone which may indicate higher actin turnover rates in these cells. Site directed mutagenesis generated several mutants for WDR1/ WDRΔ35 which indicated that specific residues (e.g. WDRR17G, WDRΔ35H48Q and WDRΔ35G204E) could alter the ratio of G/F-actin in cells, suggesting an important structural role in WDR1 function.

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Biology Commons

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