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p95-APP1 links membrane transport to Rac-mediated reorganization of actin

Nature Cell Biology volume 2, pages 521–530 (2000)Cite this article

Abstract

Motility requires protrusive activity at the cellular edge, where Rho family members regulate actin dynamics. Here we show that p95-APP1 (ArfGAP-putative, Pix-interacting, paxillin-interacting protein 1), a member of the GIT1/PKL family, is part of a complex that interacts with Rac. Wild-type and truncated p95-APP1 induce actin-rich protrusions mediated by Rac and ADP-ribosylation factor 6 (Arf6). Distinct p95-APP1-derived polypeptides have different distributions, indicating that p95-APP1 cycles between the cell surface and endosomes. Our results show that p95-APP1 functionally interacts with Rac and localizes to endosomal compartments, thus identifying p95-APP1 as a molecular link between actin organization, adhesion, and membrane transport during cell motility.

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Figure 1: Specific interaction of p95 with activated Rac.
Figure 2: Predicted amino-acid sequence of p95-APP1 and comparison to GIT1 and p95PKL.
Figure 3: Biochemical characterization of the p95-APP1–Rac interaction.
Figure 4: Distinct portions of p95-APP1 have distinct subcellular distributions and effects.
Figure 5: Differential subcellular distribution of p95-GAP and p95-N4.
Figure 6: p95-APP1 may be redistributed between the plasma membrane and the endosomal compartment.
Figure 7: PIX co-localizes with p95 and p95-C2 to a distinct intracellular compartment.
Figure 8: Cycling Arf6 is required for p95-APP1-induced cytoskeletal reorganization.
Figure 9: Rac mediates p95-C-induced protrusion and affects the subcellular distribution of p95-C.
Figure 10: Summary of the results and model for p95-APP1 function.

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Acknowledgements

We are grateful to B. Ranscht for the E13 chick-brain cDNA library, to E. Manser and L. Lim for the pXJ40–HA–βPIX plasmid and the anti-PIX antibody, to V. Hsu for the anti-Arf6 antibody, to G. Bokoch for the pCMV6m/Pak1 plasmid and the anti-Pak1 antibody, and to J. Meldolesi for critical reading of the manuscript. The financial support of Telethon-Italy (grant n.1171 to I.d.C.) is gratefully acknowledged. A.D.C. was supported by a fellowship from the Armenise-Harvard Foundation. C.A. was supported by a fellowship from the Italian Federation for Cancer Research (FIRC).

Correspondence and requests for materials should be addressed to I.d.C. The nucleotide sequence of p95-APP1 has been deposited at Genbank under accession number AF216970.

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  1. Alessandra Di Cesare, Simona Paris and Renato Longhi: These authors contributed equally to this work

Authors and Affiliations

  1. Cell Adhesion Unit - DIBIT - S. Raffaele Scientific Institute, Via Olgettina 58, Milano, 20132, Italy

    Alessandra Di Cesare, Simona Paris, Chiara Albertinazzi, Simona Dariozzi & Ivan de Curtis

  2. Center for Experimental BioInformatics, Odense University , Staermosegaardsvej 16, Odense M, DK-5230, Denmark

    Jens Andersen & Matthias Mann

  3. Institute of Biocatalysis and Molecular Recognition, National Research Council, Via Mario Bianco 9, Milano, 20133, Italy

    Renato Longhi

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Di Cesare, A., Paris, S., Albertinazzi, C. et al. p95-APP1 links membrane transport to Rac-mediated reorganization of actin . Nat Cell Biol 2, 521–530 (2000). https://doi.org/10.1038/35019561

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