Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

An EGFR/PI3K/AKT axis promotes accumulation of the Rac1-GEF Tiam1 that is critical in EGFR-driven tumorigenesis

Oncogene volume 34, pages 5971–5982 (2015)Cite this article

Subjects

Abstract

Epidermal growth factor receptor (EGFR) signaling regulates cell growth and survival, and its overactivation drives cancer development. One important branch of EGFR signaling is through activation of GTPase Rac1, which further promotes cell proliferation, survival and cancer metastasis. Here, we show that EGFR activates Rac1 via inducing the accumulation of its specific guanine nucleotide exchange factor, T-cell lymphoma invasion and metastasis 1 (Tiam1) in non-small-cell lung cancer and colon cancer cells. Conversely, elevated Tiam1 is required for EGFR-induced tumorigenesis. In human lung adenocarcinoma and colon cancer specimens, Tiam1 expression strongly correlates with EGFR expression. We further reveal that AKT, a key downstream protein kinase of EGFR, phosphorylates Tiam1 at several consensus sites, facilitates the interaction of Tiam1 with scaffold proteins 14-3-3 and leads to an increase of Tiam1 stability. Subsequently, Tiam1 is dephosporylated and destabilized by PP2A. Together, our study identifies a bidirectional (phosphorylation and dephosphorylation) regulatory mechanism controlling Tiam1 stability and provides new insights on how EGFR signaling triggers Rac1 activation and cancer development.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Hynes NE, Lane HA . ERBB receptors and cancer: the complexity of targeted inhibitors. Nat Rev Cancer 2005; 5: 341–354.

    Article  CAS  PubMed  Google Scholar 

  2. Avraham R, Yarden Y . Feedback regulation of EGFR signalling: decision making by early and delayed loops. Nat Rev Mol Cell Biol 2011; 12: 104–117.

    Article  CAS  PubMed  Google Scholar 

  3. Arai A, Jin A, Yan W, Mizuchi D, Yamamoto K, Nanki T et al. SDF-1 synergistically enhances IL-3-induced activation of the Raf-1/MEK/Erk signaling pathway through activation of Rac and its effector Pak kinases to promote hematopoiesis and chemotaxis. Cell Signal 2005; 17: 497–506.

    Article  CAS  PubMed  Google Scholar 

  4. Davis RJ . Signal transduction by the JNK group of MAP kinases. Cell 2000; 103: 239–252.

    Article  CAS  PubMed  Google Scholar 

  5. Jaffe AB, Hall A . Rho GTPases: biochemistry and biology. Annu Rev Cell Dev Biol 2005; 21: 247–269.

    Article  CAS  PubMed  Google Scholar 

  6. Schmidt A, Hall A . Guanine nucleotide exchange factors for Rho GTPases: turning on the switch. Genes Dev 2002; 16: 1587–1609.

    Article  CAS  PubMed  Google Scholar 

  7. Vega FM, Ridley AJ . Rho GTPases in cancer cell biology. FEBS Lett 2008; 582: 2093–2101.

    Article  CAS  PubMed  Google Scholar 

  8. Ellenbroek SI, Collard JG . Rho GTPases: functions and association with cancer. Clin Exp Metastasis 2007; 24: 657–672.

    Article  CAS  PubMed  Google Scholar 

  9. Wertheimer E, Gutierrez-Uzquiza A, Rosemblit C, Lopez-Haber C, Sosa MS, Kazanietz MG . Rac signaling in breast cancer: a tale of GEFs and GAPs. Cell Signal 2012; 24: 353–362.

    Article  CAS  PubMed  Google Scholar 

  10. Habets GG, Scholtes EH, Zuydgeest D, van der Kammen RA, Stam JC, Berns A et al. Identification of an invasion-inducing gene, Tiam-1, that encodes a protein with homology to GDP-GTP exchangers for Rho-like proteins. Cell 1994; 77: 537–549.

    Article  CAS  PubMed  Google Scholar 

  11. Malliri A, van der Kammen RA, Clark K, van der Valk M, Michiels F, Collard JG . Mice deficient in the Rac activator Tiam1 are resistant to Ras-induced skin tumours. Nature 2002; 417: 867–871.

    Article  CAS  PubMed  Google Scholar 

  12. Minard ME, Ellis LM, Gallick GE . Tiam1 regulates cell adhesion, migration and apoptosis in colon tumor cells. Clin Exp Metastasis 2006; 23: 301–313.

    Article  CAS  PubMed  Google Scholar 

  13. Malliri A, Rygiel TP, van der Kammen RA, Song JY, Engers R, Hurlstone AF et al. The rac activator Tiam1 is a Wnt-responsive gene that modifies intestinal tumor development. J Biol Chem 2006; 281: 543–548.

    Article  CAS  PubMed  Google Scholar 

  14. Adam L, Vadlamudi RK, McCrea P, Kumar R . Tiam1 overexpression potentiates heregulin-induced lymphoid enhancer factor-1/beta -catenin nuclear signaling in breast cancer cells by modulating the intercellular stability. J Biol Chem 2001; 276: 28443–28450.

    Article  CAS  PubMed  Google Scholar 

  15. Strumane K, Rygiel T, van der Valk M, Collard JG . Tiam1-deficiency impairs mammary tumor formation in MMTV-c-neu but not in MMTV-c-myc mice. J Cancer Res Clin Oncol 2009; 135: 69–80.

    Article  CAS  PubMed  Google Scholar 

  16. Ramis G, Thomas-Moya E, Fernandez de Mattos S, Rodriguez J, Villalonga P . EGFR inhibition in glioma cells modulates Rho signaling to inhibit cell motility and invasion and cooperates with temozolomide to reduce cell growth. PloS One 2012; 7: e38770.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Dise RS, Frey MR, Whitehead RH, Polk DB . Epidermal growth factor stimulates Rac activation through Src and phosphatidylinositol 3-kinase to promote colonic epithelial cell migration. Am J Physiol Gastrointest Liver Physiol 2008; 294: G276–G285.

    Article  CAS  PubMed  Google Scholar 

  18. Itoh RE, Kiyokawa E, Aoki K, Nishioka T, Akiyama T, Matsuda M . Phosphorylation and activation of the Rac1 and Cdc42 GEF Asef in A431 cells stimulated by EGF. J Cell Sci 2008; 121: 2635–2642.

    Article  CAS  PubMed  Google Scholar 

  19. Sordella R, Bell DW, Haber DA, Settleman J . Gefitinib-sensitizing EGFR mutations in lung cancer activate anti-apoptotic pathways. Science 2004; 305: 1163–1167.

    Article  CAS  PubMed  Google Scholar 

  20. Mulloy R, Ferrand A, Kim Y, Sordella R, Bell DW, Haber DA et al. Epidermal growth factor receptor mutants from human lung cancers exhibit enhanced catalytic activity and increased sensitivity to gefitinib. Cancer Res 2007; 67: 2325–2330.

    Article  CAS  PubMed  Google Scholar 

  21. Mukohara T, Engelman JA, Hanna NH, Yeap BY, Kobayashi S, Lindeman N et al. Differential effects of gefitinib and cetuximab on non-small-cell lung cancers bearing epidermal growth factor receptor mutations. J Natl Cancer Inst 2005; 97: 1185–1194.

    Article  CAS  PubMed  Google Scholar 

  22. Krause DS, Van Etten RA . Tyrosine kinases as targets for cancer therapy. N Engl J Med 2005; 353: 172–187.

    Article  CAS  PubMed  Google Scholar 

  23. Nicholson RI, Gee JM, Harper ME . EGFR and cancer prognosis. Eur J Cancer 2001; 37: S9–15.

    Article  CAS  PubMed  Google Scholar 

  24. Minard ME, Kim LS, Price JE, Gallick GE . The role of the guanine nucleotide exchange factor Tiam1 in cellular migration, invasion, adhesion and tumor progression. Breast Cancer Res Treat 2004; 84: 21–32.

    Article  CAS  PubMed  Google Scholar 

  25. Steiner P, Joynes C, Bassi R, Wang S, Tonra JR, Hadari YR et al. Tumor growth inhibition with cetuximab and chemotherapy in non-small cell lung cancer xenografts expressing wild-type and mutated epidermal growth factor receptor. Clin Cancer Res 2007; 13: 1540–1551.

    Article  CAS  PubMed  Google Scholar 

  26. da Cunha Santos G, Shepherd FA, Tsao MS . EGFR mutations and lung cancer. Annu Rev Pathol 2011; 6: 49–69.

    Article  PubMed  Google Scholar 

  27. Caporali S, Levati L, Starace G, Ragone G, Bonmassar E, Alvino E et al. AKT is activated in an ataxia-telangiectasia and Rad3-related-dependent manner in response to temozolomide and confers protection against drug-induced cell growth inhibition. Mol Pharmacol 2008; 74: 173–183.

    Article  CAS  PubMed  Google Scholar 

  28. Alessi DR, Caudwell FB, Andjelkovic M, Hemmings BA, Cohen P . Molecular basis for the substrate specificity of protein kinase B; comparison with MAPKAP kinase-1 and p70 S6 kinase. FEBS Lett 1996; 399: 333–338.

    Article  CAS  PubMed  Google Scholar 

  29. Hermeking H . The 14-3-3 cancer connection. Nat Rev Cancer 2003; 3: 931–943.

    Article  CAS  PubMed  Google Scholar 

  30. Fu H, Subramanian RR, Masters SC . 14-3-3 proteins: structure, function, and regulation. Annu Rev Pharmacol Toxicol 2000; 40: 617–647.

    Article  CAS  PubMed  Google Scholar 

  31. Morrison DK . The 14-3-3 proteins: integrators of diverse signaling cues that impact cell fate and cancer development. Trends Cell Biol 2009; 19: 16–23.

    Article  CAS  PubMed  Google Scholar 

  32. Woodcock SA, Jones RC, Edmondson RD, Malliri A . A modified tandem affinity purification technique identifies that 14-3-3 proteins interact with Tiam1, an interaction which controls Tiam1 stability. J Proteome Res 2009; 8: 5629–5641.

    Article  CAS  PubMed  Google Scholar 

  33. Sato S, Fujita N, Tsuruo T . Modulation of Akt kinase activity by binding to Hsp90. Proc Natl Acad Sci USA 2000; 97: 10832–10837.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Xu Y, Xing Y, Chen Y, Chao Y, Lin Z, Fan E et al. Structure of the protein phosphatase 2 A holoenzyme. Cell 2006; 127: 1239–1251.

    Article  CAS  PubMed  Google Scholar 

  35. Eichhorn PJ, Creyghton MP, Bernards R . Protein phosphatase 2 A regulatory subunits and cancer. Biochim Biophys Acta 2009; 1795: 1–15.

    CAS  PubMed  Google Scholar 

  36. Fernandez-Zapico ME, Gonzalez-Paz NC, Weiss E, Savoy DN, Molina JR, Fonseca R et al. Ectopic expression of VAV1 reveals an unexpected role in pancreatic cancer tumorigenesis. Cancer Cell 2005; 7: 39–49.

    Article  CAS  PubMed  Google Scholar 

  37. Laplante M, Sabatini DM . mTOR signaling in growth control and disease. Cell 2012; 149: 274–293.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Kumar R, Gururaj AE, Barnes CJ . p21-activated kinases in cancer. Nat Rev Cancer 2006; 6: 459–471.

    Article  CAS  PubMed  Google Scholar 

  39. Gao Y, Dickerson JB, Guo F, Zheng J, Zheng Y . Rational design and characterization of a Rac GTPase-specific small molecule inhibitor. Proc Natl Acad Sci USA 2004; 101: 7618–7623.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Shutes A, Onesto C, Picard V, Leblond B, Schweighoffer F, Der CJ . Specificity and mechanism of action of EHT 1864, a novel small molecule inhibitor of Rac family small GTPases. J Biol Chem 2007; 282: 35666–35678.

    Article  CAS  PubMed  Google Scholar 

  41. Buchanan FG, Elliot CM, Gibbs M, Exton JH . Translocation of the Rac1 guanine nucleotide exchange factor Tiam1 induced by platelet-derived growth factor and lysophosphatidic acid. J Biol Chem 2000; 275: 9742–9748.

    Article  CAS  PubMed  Google Scholar 

  42. Michiels F, Stam JC, Hordijk PL, van der Kammen RA, Ruuls-Van Stalle L, Feltkamp CA et al. Regulated membrane localization of Tiam1, mediated by the NH2-terminal pleckstrin homology domain, is required for Rac-dependent membrane ruffling and C-Jun NH2-terminal kinase activation. J Cell Biol 1997; 137: 387–398.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Viaud J, Lagarrigue F, Ramel D, Allart S, Chicanne G, Ceccato L et al. Phosphatidylinositol 5-phosphate regulates invasion through binding and activation of Tiam1. Nat Commun 2014; 5: 4080.

    Article  CAS  PubMed  Google Scholar 

  44. O'Toole TE, Bialkowska K, Li X, Fox JE . Tiam1 is recruited to beta1-integrin complexes by 14-3-3zeta where it mediates integrin-induced Rac1 activation and motility. J Cell Physiol 2011; 226: 2965–2978.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Kobayashi H, Ogura Y, Sawada M, Nakayama R, Takano K, Minato Y et al. Involvement of 14-3-3 proteins in the second epidermal growth factor-induced wave of Rac1 activation in the process of cell migration. J Biol Chem 2011; 286: 39259–39268.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Sosa MS, Lopez-Haber C, Yang C, Wang H, Lemmon MA, Busillo JM et al. Identification of the Rac-GEF P-Rex1 as an essential mediator of ErbB signaling in breast cancer. Mol Cell 2010; 40: 877–892.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Ebi H, Costa C, Faber AC, Nishtala M, Kotani H, Juric D et al. PI3K regulates MEK/ERK signaling in breast cancer via the Rac-GEF, P-Rex1. Proc Natl Acad Sci USA 2013; 110: 21124–21129.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Engers R, Zwaka TP, Gohr L, Weber A, Gerharz CD, Gabbert HE . Tiam1 mutations in human renal-cell carcinomas. Int J Cancer 2000; 88: 369–376.

    Article  CAS  PubMed  Google Scholar 

  49. Hanahan D, Weinberg RA . Hallmarks of cancer: the next generation. Cell 2011; 144: 646–674.

    Article  CAS  PubMed  Google Scholar 

  50. Vlacich G, Coffey RJ . Resistance to EGFR-targeted therapy: a family affair. Cancer cell 2011; 20: 423–425.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Yonesaka K, Zejnullahu K, Okamoto I, Satoh T, Cappuzzo F, Souglakos J et al. Activation of ERBB2 signaling causes resistance to the EGFR-directed therapeutic antibody cetuximab. Sci Transl Med 2011; 3: 99ra86.

    Article  PubMed  PubMed Central  Google Scholar 

  52. Bid HK, Roberts RD, Manchanda PK, Houghton PJ . RAC1: an emerging therapeutic option for targeting cancer angiogenesis and metastasis. Mol Cancer Ther 2013; 12: 1925–1934.

    Article  CAS  PubMed  Google Scholar 

  53. Hofbauer SW, Krenn PW, Ganghammer S, Asslaber D, Pichler U, Oberascher K et al. Tiam1/Rac1 signals contribute to the proliferation and chemoresistance, but not motility, of chronic lymphocytic leukemia cells. Blood 2014; 123: 2181–2188.

    Article  CAS  PubMed  Google Scholar 

  54. Karpel-Massler G, Westhoff MA, Zhou S, Nonnenmacher L, Dwucet A, Kast RE et al. Combined inhibition of HER1/EGFR and RAC1 results in a synergistic antiproliferative effect on established and primary cultured human glioblastoma cells. Mol Cancer Ther 2013; 12: 1783–1795.

    Article  CAS  PubMed  Google Scholar 

  55. Zhu G, Fan Z, Ding M, Mu L, Liang J, Ding Y et al. DNA damage induces the accumulation of Tiam1 by blocking beta-TrCP-dependent degradation. J Biol Chem 2014; 289: 15482–15494.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Palamidessi A, Frittoli E, Garre M, Faretta M, Mione M, Testa I et al. Endocytic trafficking of Rac is required for the spatial restriction of signaling in cell migration. Cell 2008; 134: 135–147.

    Article  CAS  PubMed  Google Scholar 

  57. Tokuzawa Y, Kaiho E, Maruyama M, Takahashi K, Mitsui K, Maeda M et al. Fbx15 is a novel target of Oct3/4 but is dispensable for embryonic stem cell self-renewal and mouse development. Mol Cell Biol 2003; 23: 2699–2708.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Su X, Zhu G, Ding X, Lee SY, Dou Y, Zhu B et al. Molecular basis underlying histone H3 lysine-arginine methylation pattern readout by Spin/Ssty repeats of Spindlin1. Genes Dev 2014; 28: 622–636.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Habas R, He X . Activation of Rho and Rac by Wnt/frizzled signaling. Methods Enzymol 2006; 406: 500–511.

    Article  CAS  PubMed  Google Scholar 

  60. Wang L, Piao T, Cao M, Qin T, Huang L, Deng H et al. Flagellar regeneration requires cytoplasmic microtubule depolymerization and kinesin-13. J Cell Sci 2013; 126: 1531–1540.

    Article  CAS  PubMed  Google Scholar 

  61. Zhu G, Wang Y, Huang B, Liang J, Ding Y, Xu A et al. A Rac1/PAK1 cascade controls beta-catenin activation in colon cancer cells. Oncogene 2012; 31: 1001–1012.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Drs Ye-Guang Chen, John G. Collard, Chuanmao Zhang, Joseph R Testa and Yigong Shi for providing plasmids and reagents. We are grateful for Dr Haiteng Deng and Jieyuan Liu for mass spectrometry analysis. This work was supported by grants to WW from the National Natural Science Foundation of China (31221064 and 81272235). GZ is a postdoctoral fellow supported by Tsinghua-Peking Center for Life Sciences.

Author information

Author notes

  1. G Zhu and Z Fan: These authors contributed equally to this work.

Authors and Affiliations

  1. MOE Key Laboratory of Protein Science, School of Life Sciences, Tsinghua University, Beijing, China

    G Zhu, Z Fan, L Mu, Y Ding & W Wu

  2. Tsinghua-Peking Center for Life Sciences, Beijing, China

    G Zhu

  3. School of Life Sciences, Peking University, Beijing, China

    M Ding

  4. National Institute of Biological Sciences, Beijing, China

    H Zhang, L Mu & L Chen

  5. Department of Basic Medical Sciences, School of Medicine, Tsinghua University, Beijing, China

    Y Zhang & Z Chang

  6. Departments of General Surgery and Pathology, Chinese PLA General Hospital, Beijing, China

    B Jia

Authors
  1. G Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Z Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. L Mu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Y Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Y Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. B Jia
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. L Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Z Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. W Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to W Wu.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhu, G., Fan, Z., Ding, M. et al. An EGFR/PI3K/AKT axis promotes accumulation of the Rac1-GEF Tiam1 that is critical in EGFR-driven tumorigenesis. Oncogene 34, 5971–5982 (2015). https://doi.org/10.1038/onc.2015.45

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2015.45

This article is cited by

Search

Advanced search

Quick links