FIG 5.
The Rho GEF or PDZ binding motifs of Map are not required for ADAM10-EGFR-MAPK activation. (A) Schematic presentation of wild-type Map (Mapwt) and Map mutants (MapWxxxA and MapΔTRL) used in this study. The Map mitochondrial targeting signal (MTS; amino acids [aa] 1 to 44), the WxxxE Rho GEF domain (aa 74 to 78), the mitochondrial toxicity region (MTR, aa 101 to 152), and the C-terminal PDZ type I binding TRL motif (aa 201 to 203) are indicated. (B and C) Effects of translocated Map on pERK and pEGFR levels. HeLa cells were infected with the indicated EPEC strains, and the levels of pERK and pEGFR in the infected cells were determined, as described in Materials and Methods. Results are means and SE of 3 independent experiments. (D) Effects of MAP on ADAM10 sheddase activity. HeLa cells transfected with the BTC-AP encoding plasmid were infected with the indicated EPEC strains for 30 min at 37°C and the level of ADAM10 sheddase activity was determined as described in Materials and Methods. (E) Map translocation. HeLa cells were infected with the indicated EPEC strains and the Map translocation into the infected cells was assessed by the effector translocation assay, as described in Materials and Methods. ***, P < 0.0005; **, P < 0.005.