Abstract
Mechanotransduction in cells describes the process by which external physical stimuli are converted into biochemical activity and plays an important role in many biological functions on both the cell and tissue level. However, the specific mechanisms by which mechanical forces lead to particular molecular and cellular responses are much less understood. We investigate the functional changes of non-erythroid α-II and β-II spectrins as a result of equi-biaxial strain application to cells in culture. Specifically, we focus on the spectrins’ role in the ubiquitination process of spectrin associated ankyrin, a vital process in the regulation of its degredation as well as its interactions with other membrane proteins. We utilize epifluorescenc, FRET and TIRF microscopy, immune-fluorescence staining and fluorescent fusion proteins for quantitative fluorescence imaging of spectrin, ankyrin and ubiquitin in live and fixed cells. Protein expression levels and localization between cells exposed to mechanical stimuli of different temporal and spatial profiles are compared. In addition, the threshold behavior of cell proliferation - as measured by number densities - of CHO, 10T1/2 and 3T3 cells as a function of mechanostimulation with different frequencies and durations is investigated in depth.Key words:Ankyrin , FRET, Proliferation, Spectrin, Ubiqutination,
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