A role for POR1, a Rac1-interacting protein, in ARF6-mediated cytoskeletal rearrangements

The ARF6 GTPase, the least conserved member of the ADP ribosylation factor (ARF) family, associates,vith the plasma membrane and intracellular endosome vesicles, Mutants of ARF6 defective in GTP binding and hydrolysis have a marked effect on endocytic trafficking and the gross morphology of the peripheral membrane system, Here we report that expression of the GTPase-defective mutant of ARF6, ARF6(Q67L), remodels the actin cytoskeleton by inducing actin polymerization at the cell periphery, This cytoskeletal rearrangement was inhibited by co-expression of ARF6(Q67L) with deletion mutants of POR1, a Rac1-interacting protein involved in membrane ruffling, but not,vith the dominant-negative mutant of Rad, Rac1(S17N), A synergistic effect between POR1 and ARF6 for the induction of actin polymerization was detected, Furthermore, we observed that ARF6 interacts directly with POR1 and that this interaction was GTP dependent, These findings indicate that ARF6 and Rad function on distinct signaling pathways to mediate cytoskeletal reorganization, and suggest a role for POR1 as an important regulatory element in orchestrating cytoskeletal rearrangements at the cell periphery induced by ARF6 and Rac1.