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IRSp53 is an essential intermediate between Rac and WAVE in the regulation of membrane ruffling

Abstract

Neural Wiskott-Aldrich syndrome protein (N-WASP)1 functions in several intracellular events including filopodium formation2, vesicle transport3 and movement of Shigella frexneri4,5 and vaccinia virus6, by stimulating rapid actin polymerization through the Arp2/3 complex. N-WASP is regulated by the direct binding of Cdc42 (refs 7, 8 ), which exposes the domain in N-WASP that activates the Arp2/3 complex2,9. A WASP-related protein, WAVE/Scar, functions in Rac-induced membrane ruffling10,11; however, Rac does not bind directly to WAVE10, raising the question of how WAVE is regulated by Rac. Here we demonstrate that IRSp53, a substrate for insulin receptor12 with unknown function, is the ‘missing link’ between Rac and WAVE. Activated Rac binds to the amino terminus of IRSp53, and carboxy-terminal Src-homology-3 domain of IRSp53 binds to WAVE to form a trimolecular complex. From studies of ectopic expression, we found that IRSp53 is essential for Rac to induce membrane ruffling, probably because it recruits WAVE, which stimulates actin polymerization mediated by the Arp2/3 complex.

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Figure 1: The specific and direct interaction of IRSp53 and WAVE2.
Figure 2: Importance of IRSp53/WAVE2 in Rac-induced membrane ruffling.
Figure 3: Direct association of activated Rac with the N-terminal region of IRSp53 (RCB).
Figure 4: The IRSp53-mediated association of Rac and WAVE2.
Figure 5: Stimulation of Arp2/3 complex activation by WAVE2 in the presence of IRSp53.

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Acknowledgements

We thank K. Miki for technical assistance in yeast two-hybrid screening and characterization of IRSp53, and N. Sasaki for WAVE1 proteins and recombinant baculoviruses. We are also grateful to T. Suzuki and C. Sasakawa for the gift of the CRIB motif fragment of PAK. This study was supported in part by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science, and Culture of Japan.

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Correspondence to Tadaomi Takenawa.

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Miki, H., Yamaguchi, H., Suetsugu, S. et al. IRSp53 is an essential intermediate between Rac and WAVE in the regulation of membrane ruffling. Nature 408, 732–735 (2000). https://doi.org/10.1038/35047107

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