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α4 Integrins are Type I cAMP-dependent protein kinase-anchoring proteins

Nature Cell Biology volume 9, pages 415–421 (2007)Cite this article

Abstract

A-kinase anchoring proteins (AKAPs) control the localization and substrate specificity of cAMP-dependent protein kinase (PKA), tetramers of regulatory (PKA-R) and catalytic (PKA-C) subunits, by binding to PKA-R subunits1. Most mammalian AKAPs bind Type II PKA through PKA-RII (ref. 2), whereas dual specificity AKAPs bind both PKA-RI and PKA-RII (ref. 3). Inhibition of PKA–AKAP interactions modulates PKA signalling2. Localized PKA activation in pseudopodia of migrating cells4 phosphorylates α4 integrins to provide spatial cues governing cell motility5. Here, we report that the α4 cytoplasmic domain is a Type I PKA-specific AKAP that is distinct from canonical AKAPs in two ways: the α4 interaction requires the PKA holoenzyme, and is insensitive to amphipathic peptides that disrupt most PKA–AKAP interactions. We exploited type-specific PKA anchoring peptides to create genetically encoded baits that sequester specific PKA isoforms to the mitochondria and found that mislocalization of Type I, but not Type II, PKA disrupts α4 phosphorylation and markedly inhibits the velocity and directional persistence of cell migration.

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Figure 1: Integrin α4β1 associates with Type I PKA.
Figure 2: Integrin α4 interacts directly with Type I PKA.
Figure 3: PKA-RIα is required for phosphorylation of α4 and other substrates at the leading edge.
Figure 4: Delocalization of Type I PKA reduces α4 phosphorylation at the leading edge and inhibits α4β1-dependent cell migration.

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Acknowledgements

We gratefully acknowledge L. Goldfinger and N. Nishiya for helpful discussions, and M. Deal for technical assistance. This work was supported by grants from the National Institutes of Health (NIH) to M.H.G., the Leukemia and Lymphoma Society to C.J.L., and Arthritis Foundation to J.H.

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Authors and Affiliations

  1. Department of Medicine, University of California San Diego, La Jolla, 92093, CA, USA

    Chinten James Lim, Jaewon Han, Nima Yousefi & Mark H. Ginsberg

  2. Department of Chemistry and Biochemistry and Howard Hughes Medical Institute, University of California San Diego, La Jolla, 92093, CA, USA

    Yuliang Ma & Susan S. Taylor

  3. Department of Pharmacology, University of Washington, Seattle, 98195, WA, USA

    Paul S. Amieux & G. Stanley McKnight

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  1. Chinten James Lim
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Contributions

C.J.L. performed all experiments with contributions from J.H., N.Y. and P.S.A. C.J.L., J.H. and M.H.G. planned and interpreted experimental data. Y.M., S.S.T., P.S.A. and G.S.K. provided critical reagents. C.J.L., M.H.G., S.S.T and G.S.K. wrote the paper.

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Correspondence to Mark H. Ginsberg.

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The authors declare no competing financial interests.

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Supplementary figures S1, S2, S3, S4, S5 and S6 (PDF 560 kb)

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Lim, C., Han, J., Yousefi, N. et al. α4 Integrins are Type I cAMP-dependent protein kinase-anchoring proteins. Nat Cell Biol 9, 415–421 (2007). https://doi.org/10.1038/ncb1561

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