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Protein kinase D1 regulates cofilin-mediated F-actin reorganization and cell motility through slingshot

Nature Cell Biology volume 11, pages 545–556 (2009)Cite this article

Abstract

Dynamic actin remodelling processes at the leading edge of migrating tumour cells are concerted events controlled by a fine-tuned temporal and spatial interplay of kinases and phosphatases. Actin severing is regulated by actin depolymerizing factor (ADF)/cofilin, which regulates stimulus-induced lamellipodia protrusion and directed cell motility. Cofilin is activated by dephosphorylation through phosphatases of the slingshot (SSH) family. SSH activity is strongly increased by its binding to filamentous actin (F-actin); however, other upstream regulators remain unknown. Here we show that in response to RhoA activation, protein kinase D1 (PKD1) phosphorylates the SSH enzyme SSH1L at a serine residue located in its actin-binding motif. This generates a 14-3-3-binding motif and blocks the localization of SSH1L to F-actin-rich structures in the lamellipodium by sequestering it in the cytoplasm. Consequently, expression of constitutively active PKD1 in invasive tumour cells enhanced the phosphorylation of cofilin and effectively blocked the formation of free actin-filament barbed ends and directed cell migration.

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Figure 1: Co-localization of PKD1 with SSH1L at peripheral F-actin structures.
Figure 2: PKD1 phosphorylates SSH1L in vitro and in vivo.
Figure 3: PKD1 phosphorylates SSH1L downstream of RhoA.
Figure 4: PKD1-mediated phosphorylation regulates SSH1L localization.
Figure 5: Phosphorylation of SSH1L at S978 controls its association with 14-3-3 proteins.
Figure 6: PKD1 regulates the phosphorylation of cofilin at S3.
Figure 7: PKD1 regulates the formation of free actin-filament barbed ends and cell migration.

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Acknowledgements

We thank L. Lewis-Tuffin for help with the video microscopy, and R. Mooney (University of Rochester Medical Center, New York) for providing MTLn3 cells. This work was sponsored in part by funds from the Mayo Foundation and the Mayo Comprehensive Cancer Center, the Mayo Clinic Breast Cancer SPORE (CA116201-03DR4), a 'Friends for an Earlier Breast Cancer Test' Grant (all to P.S.) and by a grant-in-aid for scientific research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (to K.M.).

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Authors and Affiliations

  1. Department of Cancer Biology, Mayo Clinic Comprehensive Cancer Center, Mayo Clinic, 4500 San Pablo Road, Jacksonville, 32224, Florida, USA

    Tim Eiseler, Heike Döppler, Irene K. Yan & Peter Storz

  2. Department of Biomolecular Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, Miyagi, 980-8578, Japan

    Kanae Kitatani & Kensaku Mizuno

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This publication is a collaborative work of the laboratories of P.S. and K.M. Data shown were generated by T.E., H.D., I.K.Y. and K.K. The data were evaluated by P.S. The manuscript was written by P.S. and K.M.

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Correspondence to Peter Storz.

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Eiseler, T., Döppler, H., Yan, I. et al. Protein kinase D1 regulates cofilin-mediated F-actin reorganization and cell motility through slingshot. Nat Cell Biol 11, 545–556 (2009). https://doi.org/10.1038/ncb1861

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