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JNK phosphorylates paxillin and regulates cell migration

Abstract

The c-Jun amino-terminal kinase (JNK) is generally thought to be involved in inflammation, proliferation and apoptosis1,2. Accordingly, its substrates are transcription factors and anti-apoptotic proteins2. However, JNK has also been shown to be required for Drosophila dorsal closure3,4, and MAP kinase/ERK kinase kinase 1, an upstream kinase in the JNK pathway, has been shown to be essential for cell migration5,6. Both results imply that JNK is important in cell migration. Here we show that JNK1 is required for the rapid movement of both fish keratocytes and rat bladder tumour epithelial cells (NBT-II). Moreover, JNK1 phosphorylates serine 178 on paxillin, a focal adhesion adaptor, both in vitro and in intact cells. NBT-II cells expressing the Ser 178 → Ala mutant of paxillin (PaxS178A) formed focal adhesions and exhibited the limited movement associated with such contacts in both single-cell-migration and wound-healing assays. In contrast, cells expressing wild-type paxillin moved rapidly and retained close contacts as the predominant adhesion. Expression of PaxS178A also inhibited the migration of two other cell lines. Thus, phosphorylation of paxillin by JNK seems essential for maintaining the labile adhesions required for rapid cell migration.

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Figure 1: JNK1 is required for cell migration.
Figure 2: Paxillin is phosphorylated at Ser 178 in vitro.
Figure 3: Paxillin is also phosphorylated at Ser 178 in cells.
Figure 4: Expression of PaxS178A inhibits cell migration.a, Expression of PaxS178A changes cell morphology (top panels) and inhibits cell movement (bottom panels).
Figure 5: Expression of PaxS178A promotes focal adhesion formation.

References

  1. Ip, Y. T. & Davis, R. J. Signal transduction by the c-Jun N-terminal kinase (JNK)—from inflammation to development. Curr. Opin. Cell Biol. 10, 205–219 (1998)

    Article  CAS  Google Scholar 

  2. Barr, R. K. & Bogoyevitch, M. A. The c-Jun N-terminal protein kinase family of mitogen-activated protein kinases (JNK MAPKs). Int. J. Biochem. Cell Biol. 33, 1047–1063 (2001)

    Article  CAS  Google Scholar 

  3. Riesgo-Escovar, J. R., Jenni, M., Fritz, A. & Hafen, E. The Drosophila Jun-N-terminal kinase is required for cell morphogenesis but not for DJun-dependent cell fate specification in the eye. Genes Dev. 10, 2759–2768 (1996)

    Article  CAS  Google Scholar 

  4. Sluss, H. K., Han, Z., Barrett, T., Davis, R. J. & Ip, Y. T. A JNK signal transduction pathway that mediates morphogenesis and an immune response in Drosophila. Genes Dev. 10, 2745–2758 (1996)

    Article  CAS  Google Scholar 

  5. Xia, Y. et al. MEK kinase 1 is critically required for c-Jun N-terminal kinase activation by proinflammatory stimuli and growth factor-induced cell migration. Proc. Natl Acad. Sci. USA 97, 5243–5248 (2000)

    Article  CAS  Google Scholar 

  6. Yujiri, T. et al. MEK kinase 1 gene disruption alters cell migration and c-Jun NH2-terminal kinase regulation but does not cause a measurable defect in NF-κB activation. Proc. Natl Acad. Sci. USA 97, 7272–7277 (2000)

    Article  CAS  Google Scholar 

  7. Ozanne, B. W. et al. Transcriptional regulation of cell invasion: AP-1 regulation of a multigenic invasion programme. Eur. J. Cancer 36, 1640–1648 (2000)

    Article  CAS  Google Scholar 

  8. Almeida, E. A. et al. Matrix survival signaling: From fibronectin via focal adhesion kinase to c-Jun NH2-terminal kinase. J. Cell Biol. 149, 741–754 (2000)

    Article  CAS  Google Scholar 

  9. Liu, S. et al. Binding of paxillin to α4 integrins modifies integrin-dependent biological responses. Nature 402, 676–681 (1999)

    Article  CAS  Google Scholar 

  10. Hagel, M. et al. The adaptor protein paxillin is essential for normal development in the mouse and is a critical transducer of fibronectin signaling. Mol. Cell. Biol. 22, 901–915 (2002)

    Article  CAS  Google Scholar 

  11. Schaller, M. D. Paxillin: A focal adhesion-associated adaptor protein. Oncogene 20, 6459–6472 (2001)

    Article  CAS  Google Scholar 

  12. Bellis, S. L., Perrotta, J. A., Curtis, M. S. & Turner, C. E. Adhesion of fibroblasts to fibronectin stimulates both serine and tyrosine phosphorylation of paxillin. Biochem. J. 325, 375–381 (1997)

    Article  CAS  Google Scholar 

  13. Vadlamudi, R., Adam, L., Talukder, A., Mendelsohn, J. & Kumar, R. Serine phosphorylation of paxillin by heregulin-β1: role of p38 mitogen activated protein kinase. Oncogene 18, 7253–7264 (1999)

    Article  CAS  Google Scholar 

  14. Brown, M. C., Perrotta, J. A. & Turner, C. E. Serine and threonine phosphorylation of the paxillin LIM domains regulates paxillin focal adhesion localization and cell adhesion to fibronectin. Mol. Biol. Cell. 9, 1803–1816 (1998)

    Article  CAS  Google Scholar 

  15. Beningo, K. A., Dembo, M., Kaverina, I., Small, J. V. & Wang, Y. L. Nascent focal adhesions are responsible for the generation of strong propulsive forces in migrating fibroblasts. J. Cell Biol. 153, 881–888 (2001)

    Article  CAS  Google Scholar 

  16. Lee, J. & Jacobson, K. The composition and dynamics of cell–substratum adhesions in locomoting fish keratocytes. J. Cell Sci. 110, 2833–2844 (1997)

    CAS  PubMed  Google Scholar 

  17. Abassi, Y. A. & Vuori, K. Tyrosine 221 in Crk regulates adhesion-dependent membrane localization of Crk and Rac and activation of Rac signaling. EMBO J. 21, 4571–4582 (2002)

    Article  CAS  Google Scholar 

  18. Hauck, C. R. et al. Inhibition of focal adhesion kinase expression or activity disrupts epidermal growth factor-stimulated signaling promoting the migration of invasive human carcinoma cells. Cancer Res. 61, 7079–7090 (2001)

    CAS  PubMed  Google Scholar 

  19. Huynh-Do, U. et al. Ephrin-B1 transduces signals to activate integrin-mediated migration, attachment and angiogenesis. J. Cell Sci. 115, 3073–3081 (2002)

    CAS  PubMed  Google Scholar 

  20. Ridley, A. J., Allen, W. E., Peppelenbosch, M. & Jones, G. E. Rho family proteins and cell migration. Biochem. Soc. Symp. 65, 111–123 (1999)

    CAS  PubMed  Google Scholar 

  21. Minden, A., Lin, A., Claret, F. X., Abo, A. & Karin, M. Selective activation of the JNK signaling cascade and c-Jun transcriptional activity by the small GTPases Rac and Cdc42Hs. Cell 81, 1147–1157 (1995)

    Article  CAS  Google Scholar 

  22. Christerson, L. B., Vanderbilt, C. A. & Cobb, M. H. MEKK1 interacts with alpha-actinin and localizes to stress fibers and focal adhesions. Cell Motil. Cytoskel. 43, 186–198 (1999)

    Article  CAS  Google Scholar 

  23. Fanger, G. R., Johnson, N. L. & Johnson, G. L. MEK kinases are regulated by EGF and selectively interact with Rac/Cdc42. EMBO J. 16, 4961–4972 (1997)

    Article  CAS  Google Scholar 

  24. Ilic, D. et al. Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-deficient mice. Nature 377, 539–544 (1995)

    Article  CAS  Google Scholar 

  25. Oktay, M., Wary, K. K., Dans, M., Birge, R. B. & Giancotti, F. G. Integrin-mediated activation of focal adhesion kinase is required for signaling to Jun NH2-terminal kinase and progression through the G1 phase of the cell cycle. J. Cell Biol. 145, 1461–1469 (1999)

    Article  CAS  Google Scholar 

  26. Liu, J., Huang, C. & Zhan, X. Src is required for cell migration and shape changes induced by fibroblast growth factor 1. Oncogene 18, 6700–6706 (1999)

    Article  CAS  Google Scholar 

  27. Nagata, K. et al. The MAP kinase kinase kinase MLK2 co-localizes with activated JNK along microtubules and associates with kinesin superfamily motor KIF3. EMBO J. 17, 149–158 (1998)

    Article  MathSciNet  CAS  Google Scholar 

  28. Small, J. V. & Kaverina, I. Microtubules meet substrate adhesions to arrange cell polarity. Curr. Opin. Cell. Biol. 15, 40–47 (2003)

    Article  CAS  Google Scholar 

  29. Lee, J., Ishihara, A., Oxford, G., Johnson, B. & Jacobson, K. Regulation of cell movement is mediated by stretch-activated calcium channels. Nature 400, 382–386 (1999)

    Article  CAS  Google Scholar 

  30. Huang, C., Liu, J., Haudenschild, C. C. & Zhan, X. The role of tyrosine phosphorylation of cortactin in the locomotion of endothelial cells. J. Biol. Chem. 273, 25770–25776 (1998)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank C. Raska, J. Han and R. M. Pope for their mass spectometric analysis, and K. Burridge, L. Graves and C. Otey for critically reading the manuscript. This study was supported by NIH grants to K.J. and M.D.S., the Cell Migration Consortium to K.J. and the National Institute for Dental and Cranial Research grant to K.J. and M.D.S.

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Correspondence to Ken Jacobson.

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Huang, C., Rajfur, Z., Borchers, C. et al. JNK phosphorylates paxillin and regulates cell migration. Nature 424, 219–223 (2003). https://doi.org/10.1038/nature01745

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