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Autophagic targeting of Src promotes cancer cell survival following reduced FAK signalling

Nature Cell Biology volume 14, pages 51–60 (2012)Cite this article

Abstract

Here we describe a mechanism that cancer cells use to survive when flux through the Src/FAK pathway is severely perturbed. Depletion of FAK, detachment of FAK-proficient cells or expression of non-phosphorylatable FAK proteins causes sequestration of active Src away from focal adhesions into intracellular puncta that co-stain with several autophagy regulators. Inhibition of autophagy results in restoration of active Src at peripheral adhesions, and this leads to cancer cell death. Autophagic targeting of active Src is associated with a Src–LC3B complex, and is mediated by c-Cbl. However, this is independent of c-Cbl E3 ligase activity, but is mediated by an LC3-interacting region. Thus, c-Cbl-mediated autophagic targeting of active Src can occur in cancer cells to maintain viability when flux through the integrin/Src/FAK pathway is disrupted. This exposes a previously unrecognized cancer cell vulnerability that may provide a new therapeutic opportunity.

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Figure 1: Localization of active Src is altered in the absence of FAK in SCCs.
Figure 2: Active Src co-localizes with Atg proteins.
Figure 3: Rate of autophagy is enhanced and active Src is turned over in a lysosomal-dependent manner.
Figure 4: Src activity is responsible for targeting to intracellular puncta.
Figure 5: Loss of adhesion is sufficient to promote the autophagic targeting of Src in the presence of FAK.
Figure 6: The autophagic targeting of Src prevents cancer cell death.
Figure 7: Src trafficking into intracellular puncta requires c-Cbl.
Figure 8: A c-Cbl LIR domain, but not E3 ligase activity, mediates Src autophagic targeting.

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References

  1. Thomas, S. M. & Brugge, J. S. Cellular functions regulated by Src family kinases. Annu. Rev. Cell Dev. Biol. 13, 513–609 (1997).

    Article  CAS  Google Scholar 

  2. Sandilands, E. et al. RhoB and actin polymerization coordinate Srcactivation with endosome-mediated delivery to the membrane. Dev. Cell 7, 855–869 (2004).

    Article  CAS  Google Scholar 

  3. Cartwright, C. A., Kamps, M. P., Meisler, A. I., Pipas, J. M. & Eckhart, W. pp60c-src activation in human colon carcinoma. J. Clin. Invest. 83, 2025–2033 (1989).

    Article  CAS  Google Scholar 

  4. Verbeek, B. S. et al. c-Src protein expression is increased in human breast cancer. An immunohistochemical and biochemical analysis. J. Pathol. 180, 383–388 (1996).

    Article  CAS  Google Scholar 

  5. Irby, R. B. et al. Activating SRC mutation in a subset of advanced human colon cancers. Nat. Genet 21, 187–190 (1999).

    Article  CAS  Google Scholar 

  6. Cartwright, C. A., Meisler, A. I. & Eckhart, W. Activation of the pp60c-src protein kinase is an early event in colonic carcinogenesis. Proc. Natl Acad. Sci. USA 87, 558–562 (1990).

    Article  CAS  Google Scholar 

  7. McLean, G. W. et al. Specific deletion of focal adhesion kinase suppressestumor formation and blocks malignant progression. Genes Dev. 18, 2998–3003 (2004).

    Article  CAS  Google Scholar 

  8. Morton, J. P. et al. Dasatinib inhibits the development of metastases in a mouse model of pancreatic ductal adenocarcinoma. Gastroenterology 139, 292–303 (2010).

    Article  CAS  Google Scholar 

  9. Pankiv, S. et al. p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J. Biol. Chem. 282, 24131–24145 (2007).

    Article  CAS  Google Scholar 

  10. Kirkin, V. et al. A role for NBR1 in autophagosomal degradation of ubiquitinated substrates. Mol. Cell 33, 505–516 (2009).

    Article  CAS  Google Scholar 

  11. Klionsky, D. J. et al. Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy 4, 151–175 (2008).

    Article  CAS  Google Scholar 

  12. Kimura, S., Noda, T. & Yoshimori, T. Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy 3, 452–460 (2007).

    Article  CAS  Google Scholar 

  13. Kimura, S., Fujita, N., Noda, T. & Yoshimori, T. Monitoring autophagy in mammalian cultured cells through the dynamics of LC3. Methods Enzymol. 452, 1–12 (2009).

    Article  CAS  Google Scholar 

  14. Hara, T. et al. FIP200, a ULK-interacting protein, is required for autophagosome formation in mammalian cells. J. Cell Biol. 181, 497–510 (2008).

    Article  CAS  Google Scholar 

  15. Timpson, P., Jones, G. E., Frame, M. C. & Brunton, V. G. Coordination of cell polarization and migration by the Rho family GTPases requires Src tyrosine kinase activity. Curr. Biol. 11, 1836–1846 (2001).

    Article  CAS  Google Scholar 

  16. Westhoff, M. A., Serrels, B., Fincham, V. J., Frame, M. C. & Carragher, N. O. SRC-mediated phosphorylation of focal adhesion kinase couples actin and adhesion dynamics to survival signaling. Mol. Cell Biol. 24, 8113–8133 (2004).

    Article  CAS  Google Scholar 

  17. Yokouchi, M. et al. Src-catalyzed phosphorylation of c-Cbl leads to the interdependent ubiquitination of both proteins. J. Biol. Chem. 276, 35185–35193 (2001).

    Article  CAS  Google Scholar 

  18. Andoniou, C. E., Thien, C. B. & Langdon, W. Y. Tumour induction by activated abl involves tyrosine phosphorylation of the product of the cbl oncogene. EMBO J. 13, 4515–4523 (1994).

    Article  CAS  Google Scholar 

  19. Joazeiro, C. A. et al. The tyrosine kinase negative regulator c-Cbl as a RING-type, E2-dependent ubiquitin-protein ligase. Science 286, 309–312 (1999).

    Article  CAS  Google Scholar 

  20. Levkowitz, G. et al. Ubiquitin ligase activity and tyrosine phosphorylation underlie suppression of growth factor signaling by c-Cbl/Sli-1. Mol. Cell 4, 1029–1040 (1999).

    Article  CAS  Google Scholar 

  21. Bjorkoy, G. et al. Monitoring autophagic degradation of p62/SQSTM1. Methods Enzymol. 452, 181–197 (2009).

    Article  Google Scholar 

  22. Mathew, R. et al. Autophagy suppresses tumorigenesis through elimination of p62. Cell 137, 1062–1075 (2009).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by Cancer Research UK (Program Grant to M.C.F. number: C157/A11473), a CR-UK Career Development Fellowship to S.W. and Beatson Institute core grants to O.J.S. and M.V. We thank P. Timpson (Beatson Institute, UK) for reagents.

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  1. Emma Sandilands and Bryan Serrels: These authors contributed equally to this work

Authors and Affiliations

  1. Edinburgh Cancer Research UK Centre, Institute of Genetics and Molecular Medicine, University of Edinburgh, Western General Hospital, Crewe Road South, Edinburgh EH4 2XR, UK

    Emma Sandilands, Bryan Serrels, Kenneth McLeod, Craig Stevens, Valerie G. Brunton, Simon Wilkinson & Margaret C. Frame

  2. Frankfurt Institute for Molecular Life Sciences, Goethe University, Theodor-Stern-Kai 7, Frankfurt arn Main D-60590, Germany

    David G. McEwan & Ivan Dikic

  3. Beatson Institute for Cancer Research, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK

    Jennifer P. Morton, Juan Pablo Macagno, Marcos Vidal & Owen J. Sansom

  4. School of Pathology and Laboratory Medicine, University of Western Australia, 35 Stirling Highway, Crawley, Western Australia 6009, Australia

    Wallace Y. Langdon

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  1. Emma Sandilands
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Contributions

E.S. and B.S. contributed equally to experimental work, project planning and data analysis. D.G.M., K.M., J. P. Morton and J. P. Macagno contributed to the experiments described in this manuscript. C.S., V.G.B., M.V., O.J.S. and I.D. provided intellectual input. W.Y.L. provided c-Cbl reagents. S.W. carried out electron microscopy and contributed to project planning and interpretation of data. M.C.F. was the grant holder and principal investigator under whom this work was carried out.

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Correspondence to Simon Wilkinson or Margaret C. Frame.

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The authors declare no competing financial interests.

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Sandilands, E., Serrels, B., McEwan, D. et al. Autophagic targeting of Src promotes cancer cell survival following reduced FAK signalling. Nat Cell Biol 14, 51–60 (2012). https://doi.org/10.1038/ncb2386

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