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Figure 3


Fig. 3. Specific Rac3-induced cell rounding and intracellular localization are dependent on amino acid residues 185-187. (A) Schematic representation of chimeric Rac3/Rac1 mutants used in the experiments. The asterisks indicate divergent residues. (B) Transient expression of Rac3*CAAX1 and Rac3*Cterm1 mutants in N1E-115 cells. eGFP-containing construct was co-transfected in a 1:5 ratio, to visualize transfected cells. 24 hours after transfection, cells were serum-starved for 24 hours, stained with phalloidin to visualize F-actin and photographed. Bar, 25 µm. Note that Rac3*CAAX1-induced morphology is similar to the cell rounding induced by the parental Rac3 protein, whereas the Rac3*Cterm1 mutant induces cell relaxation and spreading similar to the effect of Rac1. (C) Morphologies observed in B were scored as flat, round or protrusion bearing in two independent experiments, and presented in a bar-diagram. Control cells (ev) were transfected with pcDNA3.1 empty vector combined with eGFP-containing construct (ratio 10:1).





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