VASP and Profilin-1 Interaction in Cell Migration

Gau, David Martin (2011) VASP and Profilin-1 Interaction in Cell Migration. Undergraduate Thesis, University of Pittsburgh.

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Abstract

Profilins belong to a family of G-actin binding proteins which are thought to facilitate actinpolymerization at the leading edge of migrating cells via its polyproline interactions with majoractin nucleating and F-actin elongating proteins. The two major goals of this study are i) tospatially resolve profilins-1's (the only ubiquitously expressed member of profilin family)interaction with Ena (enabled)/VASP (vasodilator stimulated phosphoprotein) family of F-actinelongating protein, and ii) determine whether Ena/VASP regulates cell migration through itsinteraction with profilin-1. This study demonstrates the feasibility of GFP (green fluorescence protein)-based florescence resonance energy transfer (FRET) to identify profilin-1 and VASP interaction. Through acceptor photobleaching FRET (fluorescence resonance energy transfer) inMDA-MB-231 human breast cancer cells, we show that VASP and profilin-1 interaction at themembrane ruffles near the leading edge. We further show that VASP overexpression in breastcancer cells results in slower random cell motility; however VASP-induced suppression of cellmotility is partly rescued when VASP:profilin-1 interaction is downregulated These data suggestVASP utilizes profilin-1 to regulate breast cancer cell migration.

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