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Increased filamin binding to β-integrin cytoplasmic domains inhibits cell migration

Nature Cell Biology volume 3, pages 1060–1068 (2001)Cite this article

Abstract

Multicellular animal development depends on integrins. These adhesion receptors link to the actin cytoskeleton, transmitting biochemical signals and force during cell migration and interactions with the extracellular matrix. Many integrin–cytoskeleton connections are formed by filamins and talin. The β7 integrin tail binds strongly to filamin and supports less migration, fibronectin matrix assembly and focal adhesion formation than either the β1D tail, which binds strongly to talin, or the β1A tail, which binds modestly to both filamin and talin. To probe the role of filamin binding, we mapped the filamin-binding site of integrin tails and identified amino acid substitutions that led to selective loss of filamin binding to the β7 tail and gain of filamin binding to the β1A tail. These changes affected cell migration and membrane protrusions but not fibronectin matrix assembly or focal adhesion formation. Thus, tight filamin binding restricts integrin-dependent cell migration by inhibiting transient membrane protrusion and cell polarization.

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Figure 1: The β7 cytoplasmic tail inhibits cell migration and FN matrix formation, and results in peripheral localization of focal adhesions.
Figure 2: A six-amino-acid sequence in the β7 tail controls FLNa binding.
Figure 3: Valine to isoleucine substitutions in the β1A tail result in increased FLNa binding.
Figure 4: Isoleucine to valine substitutions in the β7 tail inhibit FLNa binding.
Figure 5: Valine to isoleucine substitutions in the β1A tail enhance FLNa localization and immunoprecipitation with integrins.
Figure 6: Increased FLNa binding to β-integrin tails inhibits cell migration without altering FN matrix assembly.
Figure 7: Increased FLNa binding to β-integrin tails reduces the speed of cell migration and reduces transient membrane protrusion.

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Acknowledgements

This work was supported by grants from the National Institutes of Health and The Susan G. Komen Breast Cancer Foundation. D.M.R. was supported by an advanced postdoctoral fellowship from The Juvenile Diabetes Foundation International. D.G.W was supported by a postdoctoral grant from The Arthritis Foundation. We thank X.-T. Nguyen and J. Byrnes for technical assistance.

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Authors and Affiliations

  1. Department of Vascular Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd, La Jolla, 92037, California, USA

    David A. Calderwood, William B. Kiosses, David M. Rose, Darren G. Woodside, Martin A. Schwartz & Mark H. Ginsberg

  2. Departments of Pediatrics and Pharmacology, University of Wisconsin, 1300 University Ave, Madison, 53706, Wisconsin, USA

    Anna Huttenlocher

  3. Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, 92037, California, USA

    Mark H. Ginsberg

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Correspondence to Mark H. Ginsberg.

Supplementary information

Supplementary figures

Figure S1 The β1A, β1D and β7 chimeric integrins are expressed at similar levels and mediate comparable levels of cell adhesion. (PDF 297 kb)

Figure S2 Chimeric αIIbβ3 integrins in which the β3 tail was replaced with the β1A, β1A(V36,40I) or β7(I36,40V) tail are expressed at similar levels and mediate comparable levels of cell adhesion.

Figure S3 Expression of α4β1 on Jurkat cell lines.

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Calderwood, D., Huttenlocher, A., Kiosses, W. et al. Increased filamin binding to β-integrin cytoplasmic domains inhibits cell migration. Nat Cell Biol 3, 1060–1068 (2001). https://doi.org/10.1038/ncb1201-1060

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