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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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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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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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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DOI: https://doi.org/10.1038/ncb1861
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