Article | Published:

PDK1 regulates cancer cell motility by antagonising inhibition of ROCK1 by RhoE

Nature Cell Biology volume 10, pages 127137 (2008) | Download Citation

Subjects

  • An Erratum to this article was published on 01 March 2008

Abstract

In three-dimensional matrices cancer cells move with a rounded, amoeboid morphology that is controlled by ROCK-dependent contraction of acto-myosin. In this study, we show that PDK1 is required for phosphorylation of myosin light chain and cell motility, both on deformable gels and in vivo. Depletion of PDK1 alters the localization of ROCK1 and reduces its ability to drive cortical acto-myosin contraction. This form of ROCK1 regulation does not require PDK1 kinase activity, but instead involves direct binding of PDK1 to ROCK1 at the plasma membrane; PDK1 competes directly with RhoE for binding to ROCK1. In the absence of PDK1, negative regulation by RhoE predominates, causing reduced acto-myosin contractility and motility. This work uncovers a novel non-catalytic role for PDK1 in regulating cortical acto-myosin and cell motility.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    & Cell migration in 3D matrix. Curr. Opin. Cell Biol. 17, 524–532 (2005).

  2. 2.

    , & Signaling pathways controlling primordial germ cell migration in zebrafish. J. Cell Sci. 117, 4787–4795 (2004).

  3. 3.

    , , & Reassembly of contractile actin cortex in cell blebs. J. Cell Biol. 175, 477–490 (2006).

  4. 4.

    et al. Migration of zebrafish primordial germ cells: a role for myosin contraction and cytoplasmic flow. Dev. Cell 11, 613–627 (2006).

  5. 5.

    & Blebbing of Dictyostelium cells in response to chemoattractant. Exp. Cell Res. 312, 2009–2017 (2006).

  6. 6.

    & Tumour-cell invasion and migration: diversity and escape mechanisms. Nature Rev. Cancer 3, 362–374 (2003).

  7. 7.

    & Differing modes of tumour cell invasion have distinct requirements for Rho/ROCK signalling and extracellular proteolysis. Nature Cell Biol. 5, 711–719 (2003).

  8. 8.

    , & Tumor cell migration in three dimensions. Methods Enzymol. 406, 625–643 (2006).

  9. 9.

    et al. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase). Science 273, 245–248 (1996).

  10. 10.

    & Rocks: multifunctional kinases in cell behaviour. Nature Rev. Mol. Cell Biol. 4, 446–456 (2003).

  11. 11.

    & Proteolytic and non-proteolytic migration of tumour cells and leucocytes. Biochem. Soc. Symp. 70, 277–285 (2003).

  12. 12.

    , , , & ROCK- and myosin-dependent matrix deformation enables protease-independent tumor-cell invasion in vivo. Curr. Biol. 16, 1515–1523 (2006).

  13. 13.

    et al. Induced expression of Rnd3 is associated with transformation of polarized epithelial cells by the Raf-MEK-extracellular signal-regulated kinase pathway. Mol. Cell. Biol. 20, 9364–9375 (2000).

  14. 14.

    et al. A new member of the Rho family, Rnd1, promotes disassembly of actin filament structures and loss of cell adhesion. J. Cell Biol. 141, 187–197 (1998).

  15. 15.

    et al. RhoE function is regulated by ROCK I-mediated phosphorylation. EMBO J. 24, 1170–1180 (2005).

  16. 16.

    , , , & RhoE binds to ROCK I and inhibits downstream signaling. Mol. Cell. Biol. 23, 4219–4229 (2003).

  17. 17.

    , & PDK1, the master regulator of AGC kinase signal transduction. Semin. Cell Dev. Biol. 15, 161–170 (2004).

  18. 18.

    et al. Role of vinculin in regulating focal adhesion turnover. Eur. J. Cell Biol. 85, 487–500 (2006).

  19. 19.

    et al. Paxillin phosphorylation at Ser273 localizes a GIT1–PIX–PAK complex and regulates adhesion and protrusion dynamics. J. Cell Biol. 173, 587–589 (2006).

  20. 20.

    et al. Small GTP-binding protein TC10 differentially regulates two distinct populations of filamentous actin in 3T3L1 adipocytes. Mol. Biol. Cell 13, 2334–2346 (2002).

  21. 21.

    , , , & Differential transmission of actin motion within focal adhesions. Science 315, 111–115 (2007).

  22. 22.

    , , , & Wrch-1, a novel member of the Rho gene family that is regulated by Wnt-1. Genes Dev. 15, 1796–1807 (2001).

  23. 23.

    , , , & Small GTPase Tc10 and its homologue RhoT induce N-WASP-mediated long process formation and neurite outgrowth. J. Cell Sci. 116, 155–168 (2003).

  24. 24.

    et al. Positive feedback between Dia1, LARG, and RhoA regulates cell morphology and invasion. Genes Dev. 21, 1478–1483 (2007).

  25. 25.

    Illuminating the metastatic process. Nature Rev. Cancer 7, 737–749 (2007).

  26. 26.

    , & Threonine phosphorylation of the β3 integrin cytoplasmic tail, at a site recognized by PDK1 and Akt/PKB in vitro, regulates Shc binding. J. Biol. Chem. 275, 30901–30906 (2000).

  27. 27.

    , & Cdc42-MRCK and Rho-ROCK signalling cooperate in myosin phosphorylation and cell invasion. Nature Cell Biol. 7, 255–261 (2005).

  28. 28.

    , & The Rho kinases I and II regulate different aspects of myosin II activity. J. Cell Biol. 170, 443–453 (2005).

  29. 29.

    et al. Conditional ROCK activation in vivo induces tumor cell dissemination and angiogenesis. Cancer Res. 64, 8994–9001 (2004).

  30. 30.

    , & Dynamics of RhoA and ROKα translocation in single living cells. Cell Biochem. Biophys. 45, 243–254 (2006).

  31. 31.

    , , & The p160 RhoA-binding kinase ROKα is a member of a kinase family and is involved in the reorganization of the cytoskeleton. Mol. Cell. Biol. 16, 5313–5327 (1996).

  32. 32.

    et al. Roles of PDK-1 and PKN in regulating cell migration and cortical actin formation of PTEN-knockout cells. Oncogene 23, 9348–9358 (2004).

  33. 33.

    et al. Essential role of PDK1 in regulating endothelial cell migration. J. Cell Biol. 176, 1035–1047 (2007).

  34. 34.

    et al. 3-phosphoinositide-dependent protein kinase-1 (PDK1) promotes invasion and activation of matrix metalloproteinases. BMC Cancer 6, 77 (2006).

  35. 35.

    et al. Phosphoinositide-dependent kinase 1 and p21-activated protein kinase mediate reactive oxygen species-dependent regulation of platelet-derived growth factor-induced smooth muscle cell migration. Circ. Res. 94, 1219–1226 (2004).

  36. 36.

    , , & Genomic analysis of metastasis reveals an essential role for RhoC. Nature 406, 532–535 (2000).

  37. 37.

    , , , & Rho GTPases in human breast tumours: expression and mutation analyses and correlation with clinical parameters. Br. J. Cancer 87, 635–644 (2002).

  38. 38.

    et al. An essential part for Rho-associated kinase in the transcellular invasion of tumor cells. Nature Med. 5, 221–225 (1999).

  39. 39.

    & RHO-GTPases and cancer. Nature Rev. Cancer 2, 133–142 (2002).

  40. 40.

    et al. Identification and testing of a gene expression signature of invasive carcinoma cells within primary mammary tumors. Cancer Res. 64, 8585–8594 (2004).

  41. 41.

    et al. Rho-associated kinase, a novel serine/threonine kinase, as a putative target for small GTP binding protein Rho. EMBO J. 15, 2208–2216 (1996).

  42. 42.

    et al. The small GTP-binding protein Rho binds to and activates a 160 kDa Ser/Thr protein kinase homologous to myotonic dystrophy kinase. EMBO J. 15, 1885–1893 (1996).

Download references

Acknowledgements

We thank Chris Marshall, Michael Way and lab members for their comments, members of the Biological Resources and Light Microscopy units for technical assistance and Cancer Research UK for funding.

Author information

Affiliations

  1. Tumour Cell Biology Laboratory, Cancer Research UK, London Research Institute, 44 Lincoln's Inn Fields, London, WC2A 3PX, UK.

    • Sophie Pinner
    •  & Erik Sahai

Authors

  1. Search for Sophie Pinner in:

  2. Search for Erik Sahai in:

Corresponding author

Correspondence to Erik Sahai.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Figures S1, S2, s3, S4, S5, S6, S7 and S8 and Supplementary Material

Videos

  1. 1.

    Supplementary Information

    Supplementary Movie 1

  2. 2.

    Supplementary Information

    Supplementary Movie 2

  3. 3.

    Supplementary Information

    Supplementary Movie 3

  4. 4.

    Supplementary Information

    Supplementary Movie 4

  5. 5.

    Supplementary Information

    Supplementary Movie 5

  6. 6.

    Supplementary Information

    Supplementary Movie 6

  7. 7.

    Supplementary Information

    Supplementary Movie 7

  8. 8.

    Supplementary Information

    Supplementary Movie 8

  9. 9.

    Supplementary Information

    Supplementary Movie 9

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ncb1675

Further reading