Article | Published:

STRIPAK components determine mode of cancer cell migration and metastasis

Nature Cell Biology volume 17, pages 6880 (2015) | Download Citation


The contractile actomyosin cytoskeleton and its connection to the plasma membrane are critical for control of cell shape and migration. We identify three STRIPAK complex components, FAM40A, FAM40B and STRN3, as regulators of the actomyosin cortex. We show that FAM40A negatively regulates the MST3 and MST4 kinases, which promote the co-localization of the contractile actomyosin machinery with the Ezrin/Radixin/Moesin family proteins by phosphorylating the inhibitors of PPP1CB, PPP1R14A–D. Using computational modelling, in vitro cell migration assays and in vivo breast cancer metastasis assays we demonstrate that co-localization of contractile activity and actin–plasma membrane linkage reduces cell speed on planar surfaces, but favours migration in confined environments similar to those observed in vivo. We further show that FAM40B mutations found in human tumours uncouple it from PP2A and enable it to drive a contractile phenotype, which may underlie its role in human cancer.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    & Collective cell migration in morphogenesis, regeneration and cancer. Nat. Rev. Mol. Cell Biol. 10, 445–457 (2009).

  2. 2.

    & The basics of epithelial-mesenchymal transition. J. Clin. Invest. 119, 1420–1428 (2009).

  3. 3.

    , , , & Intracellular fluid flow in rapidly moving cells. Nat. Cell Biol. 11, 1219–1224 (2009).

  4. 4.

    & Blebs lead the way: how to migrate without lamellipodia. Nat. Rev. Mol. Cell Biol. 9, 730–736 (2008).

  5. 5.

    & Mechanical modes of ‘amoeboid’ cell migration. Curr. Opin. Cell Biol. 21, 636–644 (2009).

  6. 6.

    , , & Non-muscle myosin II takes centre stage in cell adhesion and migration. Nat. Rev. Mol. Cell Biol. 10, 778–790 (2009).

  7. 7.

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

  8. 8.

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

  9. 9.

    & The plasticity of cytoskeletal dynamics underlying neoplastic cell migration. Curr. Opin. Cell Biol. 22, 690–696 (2010).

  10. 10.

    & Cancer dissemination—lessons from leukocytes. Dev. Cell 19, 13–26 (2010).

  11. 11.

    & Plasticity of cell migration: a multiscale tuning model. J. Cell Biol. 188, 11–19 (2010).

  12. 12.

    et al. Rac activation and inactivation control plasticity of tumor cell movement. Cell 135, 510–523 (2008).

  13. 13.

    & Myosin phosphatase target subunit: many roles in cell function. Biochem. Biophys. Res. Commun. 369, 149–156 (2008).

  14. 14.

    , & Role of protein phosphatase type 1 in contractile functions: myosin phosphatase. J. Biol. Chem. 279, 37211–37214 (2004).

  15. 15.

    , , & The myosin phosphatase targeting protein (MYPT) family: a regulated mechanism for achieving substrate specificity of the catalytic subunit of protein phosphatase type 1δ. Arch. Biochem. Biophys. 510, 147–159 (2011).

  16. 16.

    , & Cell adhesion: integrating cytoskeletal dynamics and cellular tension. Nat. Rev. Mol. Cell Biol. 11, 633–643 (2010).

  17. 17.

    , & Organizing the cell cortex: the role of ERM proteins. Nat. Rev. Mol. Cell Biol. 11, 276–287 (2010).

  18. 18.

    et al. Association of the myosin-binding subunit of myosin phosphatase and moesin: dual regulation of moesin phosphorylation by Rho-associated kinase and myosin phosphatase. J. Cell Biol. 141, 409–418 (1998).

  19. 19.

    & Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Biochem. J. 353, 417–439 (2001).

  20. 20.

    PP2A in the regulation of cell motility and invasion. Curr. Protein Pept. Sci. 12, 3–11 (2011).

  21. 21.

    , , , & Role of serine-threonine phosphoprotein phosphatases in smooth muscle contractility. Am. J. Physiol. Cell Physiol. 304, C485-504 (2013).

  22. 22.

    et al. Identification and characterization of a set of conserved and new regulators of cytoskeletal organization, cell morphology and migration. BMC Biol. 9, 54 (2011).

  23. 23.

    et al. CKA, a novel multidomain protein, regulates the JUN N-terminal kinase signal transduction pathway in Drosophila. Mol. Cell. Biol. 22, 1792–1803 (2002).

  24. 24.

    et al. A PP2A phosphatase high density interaction network identifies a novel striatin-interacting phosphatase and kinase complex linked to the cerebral cavernous malformation 3 (CCM3) protein. Mol. Cell. Proteomics 8, 157–171 (2009).

  25. 25.

    , , & An integrated workflow for charting the human interaction proteome: insights into the PP2A system. Mol. Syst. Biol. 5, 237 (2009).

  26. 26.

    et al. Combined functional genomic and proteomic approaches identify a PP2A complex as a negative regulator of Hippo signaling. Mol. Cell 39, 521–534 (2010).

  27. 27.

    et al. Structural probing of a protein phosphatase 2A network by chemical cross-linking and mass spectrometry. Science 337, 1348–1352 (2012).

  28. 28.

    & STRIPAK complexes: structure, biological function, and involvement in human diseases. Int. J. Biochem. Cell Biol. 47, 118–148 (2014).

  29. 29.

    et al. YSK1 is activated by the Golgi matrix protein GM130 and plays a role in cell migration through its substrate 14-3-3zeta. J. Cell Biol. 164, 1009–1020 (2004).

  30. 30.

    et al. Inhibition of cell migration by autophosphorylated mammalian sterile 20-like kinase 3 (MST3) involves paxillin and protein-tyrosine phosphatase-PEST. J. Biol. Chem. 281, 38405–38417 (2006).

  31. 31.

    et al. Reelin and stk25 have opposing roles in neuronal polarization and dendritic Golgi deployment. Cell 143, 826–836 (2010).

  32. 32.

    et al. Mst4 and Ezrin induce brush borders downstream of the Lkb1/Strad/Mo25 polarization complex. Dev. Cell 16, 551–562 (2009).

  33. 33.

    et al. Rap2A links intestinal cell polarity to brush border formation. Nat. Cell Biol. 14, 793–801 (2012).

  34. 34.

    , & SOcK, MiSTs, MASK and STicKs: the GCKIII (germinal centre kinase III) kinases and their heterologous protein-protein interactions. Biochem. J. 454, 13–30 (2013).

  35. 35.

    , & Slow border cells, a locus required for a developmentally regulated cell migration during oogenesis, encodes Drosophila C/EBP. Cell 71, 51–62 (1992).

  36. 36.

    Border-cell migration: the race is on. Nat. Rev. Mol. Cell Biol. 4, 13–24 (2003).

  37. 37.

    , & Group choreography: mechanisms orchestrating the collective movement of border cells. Nat. Rev. Mol. Cell Biol. 13, 631–645 (2012).

  38. 38.

    & Paracrine signaling through the JAK/STAT pathway activates invasive behavior of ovarian epithelial cells in Drosophila. Cell 107, 831–841 (2001).

  39. 39.

    & Guidance of cell migration by EGF receptor signaling during Drosophila oogenesis. Science 291, 131–133 (2001).

  40. 40.

    , , , & Guidance of cell migration by the Drosophila PDGF/VEGF receptor. Cell 107, 17–26 (2001).

  41. 41.

    & Evidence for tension-based regulation of Drosophila MAL and SRF during invasive cell migration. Dev. Cell 7, 85–93 (2004).

  42. 42.

    et al. Structure-function analysis of core STRIPAK proteins: a signaling complex implicated in Golgi polarization. J. Biol. Chem. 286, 25065–25075 (2011).

  43. 43.

    , & Rho-kinase/ROCK: a key regulator of the cytoskeleton and cell polarity. Cytoskeleton 67, 545–554 (2010).

  44. 44.

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

  45. 45.

    et al. Matrix geometry determines optimal cancer cell migration strategy and modulates response to interventions. Nat. Cell Biol. 15, 751–762 (2013).

  46. 46.

    , & Implementing an online tool for genome-wide validation of survival-associated biomarkers in ovarian-cancer using microarray data from 1287 patients. Endocr. Relat. Cancer 19, 197–208 (2012).

  47. 47.

    et al. Adaptor protein cerebral cavernous malformation 3 (CCM3) mediates phosphorylation of the cytoskeletal proteins ezrin/radixin/moesin by mammalian Ste20-4 to protect cells from oxidative stress. J. Biol. Chem. 287, 11556–11565 (2012).

  48. 48.

    Regulation of cellular protein phosphatase-1 (PP1) by phosphorylation of the CPI-17 family, C-kinase-activated PP1 inhibitors. J. Biol. Chem. 284, 35273–35277 (2009).

  49. 49.

    et al. Compensation mechanism in tumor cell migration: mesenchymal-amoeboid transition after blocking of pericellular proteolysis. J. Cell Biol. 160, 267–277 (2003).

  50. 50.

    et al. Calpain 2 and Src dependence distinguishes mesenchymal and amoeboid modes of tumour cell invasion: a link to integrin function. Oncogene 25, 5726–5740 (2006).

  51. 51.

    et al. Functional analyses of human and zebrafish 18-amino acid in-frame deletion pave the way for domain mapping of the cerebral cavernous malformation 3 protein. Hum. Mutat. 30, 1003–1011 (2009).

  52. 52.

    et al. CCM3 signaling through sterile 20-like kinases plays an essential role during zebrafish cardiovascular development and cerebral cavernous malformations. J. Clin. Invest. 120, 2795–2804 (2010).

  53. 53.

    et al. CCM1-ICAP-1 complex controls β1 integrin-dependent endothelial contractility and fibronectin remodeling. J. Cell Biol. 202, 545–561 (2013).

  54. 54.

    et al. Cumulative haploinsufficiency and triplosensitivity drive aneuploidy patterns and shape the cancer genome. Cell 155, 948–962 (2013).

  55. 55.

    , , , & An ezrin-rich, rigid uropod-like structure directs movement of amoeboid blebbing cells. J. Cell Sci. 124, 1256–1267 (2011).

  56. 56.

    , & Generation of compartmentalized pressure by a nuclear piston governs cell motility in a 3D matrix. Science 345, 1062–1065 (2014).

  57. 57.

    et al. A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. Nature 448, 151–156 (2007).

  58. 58.

    et al. In vivo fluorescence resonance energy transfer imaging reveals differential activation of Rho-family GTPases in glioblastoma cell invasion. J. Cell Sci. 125, 858–868 (2012).

  59. 59.

    et al. Effects of substrate stiffness on cell morphology, cytoskeletal structure, and adhesion. Cell Motil. Cytoskeleton 60, 24–34 (2005).

  60. 60.

    et al. An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1,809 patients. Breast Cancer Res. Treat. 123, 725–731 (2010).

  61. 61.

    , , , & GOBO: gene expression-based outcome for breast cancer online. PLoS ONE 6, e17911 (2011).

  62. 62.

    et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. Cancer Discov. 2, 401–404 (2012).

  63. 63.

    et al. Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci. Signal. 6, pl1 (2013).

Download references


C.D.M., S.H., M.T., A.B., G.F., P.A.B., B.T. and E.S. are financially supported by Cancer Research UK. C.D.M. was further supported by a FEBS long-term fellowship. We thank laboratory members for help and advice throughout this work. We thank N. O’Reilly and S. Kjaer for help with peptide synthesis and protein purification, and members of the BRU for help with metastasis assays.

Author information


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

    • Chris D. Madsen
    • , Steven Hooper
    •  & Erik Sahai
  2. Biotech Research and Innovation Centre (BRIC), University of Copenhagen, Ole Maaløes Vej 5 2200 Copenhagen N, Denmark

    • Chris D. Madsen
    •  & Janine T. Erler
  3. Biomolecular Modelling Laboratory, Cancer Research UK London Research Institute, 44 Lincoln’s Inn Fields London WC2A 3LY, UK

    • Melda Tozluoglu
    •  & Paul A. Bates
  4. Lymphocyte Interaction Laboratory, Cancer Research UK London Research Institute, 44 Lincoln’s Inn Fields London WC2A 3LY, UK

    • Andreas Bruckbauer
  5. Epithelial Biology Laboratory, Cancer Research UK London Research Institute, 44 Lincoln’s Inn Fields London, WC2A 3LY, UK

    • Georgina Fletcher
    •  & Barry Thompson


  1. Search for Chris D. Madsen in:

  2. Search for Steven Hooper in:

  3. Search for Melda Tozluoglu in:

  4. Search for Andreas Bruckbauer in:

  5. Search for Georgina Fletcher in:

  6. Search for Janine T. Erler in:

  7. Search for Paul A. Bates in:

  8. Search for Barry Thompson in:

  9. Search for Erik Sahai in:


C.D.M. and E.S. carried out all experiments except those noted otherwise. S.H. performed all kinase assays and immunoprecipitation assays. G.F. and B.T. performed the fly screen. M.T. and P.A.B. made the mathematical model. A.B. helped with structured illumination microscopy. C.D.M. and E.S. conceived the study and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Erik Sahai.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Information

Excel files

  1. 1.

    Supplementary Table 1: Information of the siRNA screen's.

    Sheet 1. Fly screen. List of genes that were depleted in the fly screen. Sheet 2. Fly genes and human homologous. Genes with similarities are also included. Sheet 3. siRNA sequences and catalogue number of all siRNA used in the screen. All siRNA's were purchased by Dharmacon. Sheet 4. Fly and human gene names, fly and A431 phenotypes.

  2. 2.

    Supplementary Table 2: Peptide kinase screen.

    Peptide sequences and specific phospho site are shown. Raw data from each experiment is shown.

  3. 3.

    Supplementary Table 3: qPCR primers and siRNA oligo's.

    qPCR primer sequences and siRNAs used in the study including catalog numbers.

  4. 4.

    Supplementary Table 4: Expression vectors.

    All expression vectors used in this study are shown.

  5. 5.

    Supplementary Table 5: Antibodies.

    All antibodies used in this study including provider, catalog numbers and dilutions are shown.


  1. 1.

    3D morphologies of siRNA depleted A431 cells.

    3D reconstruction of confocal stacks taken of siRNA transfected A431 cells stained for F-actin (red) and pS19-MLC (green). The cells were plated on top of collagen-1/matrigels. The movie includes siCtr, siFAM40A, siFAM40B and STRN3 depleted A431 cells sequentially.

  2. 2.

    Spatiotemporal regulation of MST3-GFP.

    Confocal time lapse movie of siRNA transfected A431-MST3-GFP cells. The cells have been serum starved for 24 h and then stimulated with FBS. Imaging is then initiated immediately and frames are taken every 20 s. When cells were treated with ROCK inhibitor (Y27632) the drug was added during serum starvation. The movie includes siCtr, ROCK inhibitor (Y27632) treated, and siCCM3 depleted A431-MST3-GFP cells sequentially.

  3. 3.

    Time-lapse movie of siRNA depleted MDA-MD231 cells on hard surfaces.

    Phase contrast time lapse movie of siRNA transfected MDA-MB-231 cells plated on a 2D planar surface. Images were taken every 5 min. The movie includes siCtr, siFAM40A, siFAM40B and siMST3&4 depleted MDA-MB-231 cells sequentially.

  4. 4.

    Time-lapse movie of siRNA depleted MDA-MD231 cells on soft surfaces.

    Phase contrast time lapse movie of siRNA transfected MDA-MB-231 cells plated on top of collagen-I/matrigels. Images were taken every 5 min. The movie includes siCtr, siFAM40A, siFAM40B and siMST3&4 depleted MDA-MB-231 cells sequentially.

About this article

Publication history





Further reading