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PtdIns(3,4,5)P3 binding is necessary for WAVE2-induced formation of lamellipodia

Nature Cell Biology volume 6, pages 420–426 (2004)Cite this article

Abstract

Polarized cell movement is triggered by the development of a PtdIns(3,4,5)P3 gradient at the membrane, which is followed by rearrangement of the actin cytoskeleton. The WASP family verprolin homologous protein (WAVE) is essential for lamellipodium formation at the leading edge by activating the Arp2/3 complex downstream of Rac GTPase. Here, we report that WAVE2 binds to PtdIns(3,4,5)P3 through its basic domain. The amino-terminal portion of WAVE2, which includes the PtdIns(3,4,5)P3-binding sequence, was localized at the leading edge of lamellipodia induced by an active form of Rac (RacDA) or by treatment with platelet-derived growth factor (PDGF). Production of PtdIns(3,4,5)P3 at the cell membrane by myristoylated phosphatidylinositol-3-OH kinase (PI(3)K) is sufficient to recruit WAVE2 in the presence of dominant-negative Rac and latrunculin, demonstrating that PtdIns(3,4,5)P3 alone is able to recruit WAVE2. Expression of a full-length mutant of WAVE2 that lacks the lipid-binding activity inhibited proper formation of lamellipodia induced by RacDA. These results suggest that one of the products of PI(3)K, PtdIns(3,4,5)P3, recruits WAVE2 to the polarized membrane and that this recruitment is essential for lamellipodium formation at the leading edge.

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Figure 1: Five amino-acid residues in the Basic1 region of WAVE2 are required for binding to phosphoinositides.
Figure 2: WAVE2-N is localized at Rac-induced lamellipodia and colocalizes with Myr-p110α in the presence of RacDN.
Figure 3: Localization of WAVE2 with PtdIns(4,5)P2 or PtdIns(3,4,5)P3.
Figure 4: Expression of the full-length WAVE2 mutant lacking phosphoinositide-binding activity inhibits proper formation of lamellipodia.

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Acknowledgements

We thank M. Kasuga for providing myristoylated PI(3)K-p110α, and M. Furutani and K. Tsujita for discussions and the PLCδ1PH constructs. This work was supported by grants-in-aid from the Ministry of Education, Culture, Sports, and Technology of Japan, and from the Japan Science and Technology Corporation.

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Authors and Affiliations

  1. Department of Biochemistry, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan

    Tsukasa Oikawa, Toshiki Itoh, Masayoshi Kato, Takeshi Ijuin, Daisuke Yamazaki, Shiro Suetsugu & Tadaomi Takenawa

  2. Division of Cancer Genomics, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan

    Hideki Yamaguchi

  3. CREST, Japan Science and Technology Corporation (JST), Minato-ku, Tokyo, 108-8639, Japan

    Tsukasa Oikawa, Toshiki Itoh, Masayoshi Kato, Takeshi Ijuin, Daisuke Yamazaki, Shiro Suetsugu & Tadaomi Takenawa

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Correspondence to Tadaomi Takenawa.

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Oikawa, T., Yamaguchi, H., Itoh, T. et al. PtdIns(3,4,5)P3 binding is necessary for WAVE2-induced formation of lamellipodia. Nat Cell Biol 6, 420–426 (2004). https://doi.org/10.1038/ncb1125

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