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Filamin is essential in actin cytoskeletal assembly mediated by p21-activated kinase 1

Nature Cell Biology volume 4, pages 681–690 (2002)Cite this article

Abstract

The serine/threonine kinase p21-activated kinase 1 (Pak1) controls the actin cytoskeletal and ruffle formation through mechanisms that are independent of GTPase activity. Here we identify filamin FLNa as a Pak1-interacting protein through a yeast two-hybrid screen using the amino terminus of Pak1 as a bait. FLNa is stimulated by physiological signalling molecules to undergo phosphorylation by Pak1 and to interact and colocalize with endogenous Pak1 in membrane ruffles. The ruffle-forming activity of Pak1 is functional in FLNa-expressing cells but not in FLNa-deficient cells. In FLNa, the Pak1-binding site involves tandem repeat 23 in the carboxyl terminus and phosphorylation takes place on serine 2152. The FLNa-binding site in Pak1 is localized between amino acids 52 and 132 in the conserved Cdc42/Rac-interacting (CRIB) domain; accordingly, FLNa binding to the CRIB domain stimulates Pak1 kinase activity. Our results indicate that FLNa may be essential for Pak1-induced cytoskeletal reorganization and that the two-way regulatory interaction between Pak1 and FLNa may contribute to the local stimulation of Pak1 activity and its targets in cytoskeletal structures.

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Figure 1: Identification of FLNa as a Pak1-binding protein.
Figure 2: Physiological signals induce Pak1 and FLNa colocalization in ruffles.
Figure 3: FLNa is substrate of Pak1.
Figure 4: Pak1-mediated ruffle formation requires FLNa.
Figure 5: FLNa is required for kinase-active Pak1-induced ruffle formation.
Figure 6: Location of the interaction domains of Pak1 and FLNa.
Figure 7: FLNa regulation of Pak1 activity.

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Acknowledgements

This study was supported by grants from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Molecular and Cellular Oncology, The University of Texas M. D. Anderson Cancer Center, Houston, 77030, Texas, USA

    Ratna K. Vadlamudi, Feng Li, Liana Adam, Diep Nguyen & Rakesh Kumar

  2. Brigham and Women's Hospital, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Yasutaka Ohta & Thomas P. Stossel

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Correspondence to Rakesh Kumar.

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Vadlamudi, R., Li, F., Adam, L. et al. Filamin is essential in actin cytoskeletal assembly mediated by p21-activated kinase 1. Nat Cell Biol 4, 681–690 (2002). https://doi.org/10.1038/ncb838

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