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Figure 6


Fig. 6. Centrosome separation is blocked by the wild-type cortactin C-terminus but not by the cortactin C-terminus containing mutated Tyr residues. (A) Wild-type cortactin and cortactin C-terminus mutants. N-terminus, amino acids (aa) 1-329; C-terminus, aa 330-546; CT(Y/F), aa 330-546 with Phe substitutions for Tyr421, Tyr466, Tyr475 and Tyr482. (B) HeLa cells transfected with GFP-CT or GFP-CT(Y/F). Arrowheads indicate the centrosomes. The insertions are the enlarged views of the centrosomal area. Scale bars: 10 µm. (C) Illustration of the dominant-negative effect the cortactin C-terminal domains exert on centrosome separation. (D) Inhibition of centrosome separation brought upon by the Myc-tagged C-terminus of wild-type cortactin (CT) but not the cortactin C-terminus containing mutated Tyr residues [CT(Y/F)]. The centrosome-separation ratio (d/D) was measured from 100 Myc-positive cells and plotted as described for Fig. 4. (E) Average centrosome separation 10 hours after thymidine-synchronization release in HeLa cells expressing Myc-tagged C-terminus of wild-type cortactin (CT) C-terminus containing mutated Tyr residues [CT(Y/F)]. The distances of two centrosomes (d) and corresponding nuclear diameters (D) of 100 cells were averaged for each group of cells.





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