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  • Original Article
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Met-driven invasive growth involves transcriptional regulation of Arhgap12

Oncogene volume 27, pages 5590–5598 (2008)Cite this article

Abstract

Invasive growth is a complex biological program triggered by hepatocyte growth factor (HGF) through its tyrosine kinase receptor encoded by the Met proto-oncogene. The program involves—besides proliferation—cell dissociation, motility and invasiveness, controlled by intracellular signals impinging on PI3K and on the small G-proteins of the Rac/Rho family. The mechanism(s) unbalancing Rac/Rho activation are still not completely clarified. Here, we describe a functional link between HGF and Arhgap12, a gene encoding a previously uncharacterized protein of the RhoGAP family. We identified Arhgap12 as a transcriptional target of HGF, through a novel gene trapping strategy. We found that Arhgap12 mRNA and protein are robustly suppressed by HGF treatment, but not by serum. Arhgap12 displayed GTPase activating protein (GAP) activity towards Rac1 and, upon overexpression, impaired cell scattering, invasion and adhesion to fibronectin in response to HGF. Consistently, Arhgap12 silencing by RNA interference selectively increased the scatter and adhesion responses. These data show that HGF-driven invasive growth involves transcriptional regulation of a Rac1-specific GAP.

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References

  • Bar-Sagi D, Hall A . (2000). Ras and Rho GTPases: a family reunion. Cell 103: 227–238.

    Article  CAS  PubMed  Google Scholar 

  • Bertotti A, Comoglio PM . (2003). Tyrosine kinase signal specificity: lessons from the HGF receptor. Trends Biochem Sci 28: 527–533.

    Article  CAS  PubMed  Google Scholar 

  • Birchmeier C, Birchmeier W, Gherardi E, Vande Woude GF . (2003). Met, metastasis, motility and more. Nat Rev Mol Cell Biol 4: 915–925.

    Article  CAS  PubMed  Google Scholar 

  • Boccaccio C, Ando M, Tamagnone L, Bardelli A, Michieli P, Battistini C et al. (1998). Induction of epithelial tubules by growth factor HGF depends on the STAT pathway. Nature 391: 285–288.

    Article  CAS  PubMed  Google Scholar 

  • Bosse T, Ehinger J, Czuchra A, Benesch S, Steffen A, Wu X et al. (2007). Cdc42 and phosphoinositide 3-kinase drive Rac-mediated actin polymerization downstream of c-Met in distinct and common pathways. Mol Cell Biol 27: 6615–6628.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Braga VM, Machesky LM, Hall A, Hotchin NA . (1997). The small GTPases Rho and Rac are required for the establishment of cadherin-dependent cell–cell contacts. J Cell Biol 137: 1421–1431.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Brummelkamp TR, Bernards R, Agami R . (2002). A system for stable expression of short interfering RNAs in mammalian cells. Science 296: 550–553.

    Article  CAS  PubMed  Google Scholar 

  • Cicchetti P, Ridley AJ, Zheng Y, Cerione RA, Baltimore D . (1995). 3BP-1, an SH3 domain binding protein, has GAP activity for Rac and inhibits growth factor-induced membrane ruffling in fibroblasts. EMBO J 14: 3127–3135.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Comoglio PM, Trusolino L . (2002). Invasive growth: from development to metastasis. J Clin Invest 109: 857–862.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D et al. (1998). A third-generation lentivirus vector with a conditional packaging system. J Virol 72: 8463–8471.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Finkielstein CV, Overduin M, Capelluto DG . (2006). Cell migration and signaling specificity is determined by the phosphatidylserine recognition motif of Rac1. J Biol Chem 281: 27317–27326.

    Article  CAS  PubMed  Google Scholar 

  • Follenzi A, Ailles LE, Bakovic S, Geuna M, Naldini L . (2000). Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences. Nat Genet 25: 217–222.

    Article  CAS  PubMed  Google Scholar 

  • Furge KA, Zhang YW, Vande Woude GF . (2000). Met receptor tyrosine kinase: enhanced signaling through adapter proteins. Oncogene 19: 5582–5589.

    Article  CAS  PubMed  Google Scholar 

  • Furukawa Y, Kawasoe T, Daigo Y, Nishiwaki T, Ishiguro H, Takahashi M et al. (2001). Isolation of a novel human gene, ARHGAP9, encoding a rho-GTPase activating protein. Biochem Biophys Res Commun 284: 643–649.

    Article  CAS  PubMed  Google Scholar 

  • Grisendi S, Chambraud B, Gout I, Comoglio PM, Crepaldi T . (2001). Ligand-regulated binding of FAP68 to the hepatocyte growth factor receptor. J Biol Chem 276: 46632–46638.

    Article  CAS  PubMed  Google Scholar 

  • Gual P, Giordano S, Williams TA, Rocchi S, Van OE, Comoglio PM . (2000). Sustained recruitment of phospholipase C-gamma to Gab1 is required for HGF-induced branching tubulogenesis. Oncogene 19: 1509–1518.

    Article  CAS  PubMed  Google Scholar 

  • Guo F, Debidda M, Yang L, Williams DA, Zheng Y . (2006). Genetic deletion of Rac1 GTPase reveals its critical role in actin stress fiber formation and focal adhesion complex assembly. J Biol Chem 281: 18652–18659.

    Article  CAS  PubMed  Google Scholar 

  • Hall A . (1998). Rho GTPases and the actin cytoskeleton. Science 279: 509–514.

    Article  CAS  PubMed  Google Scholar 

  • Hordijk PL, ten Klooster JP, van der Kammen RA, Michiels F, Oomen LC, Collard JG . (1997). Inhibition of invasion of epithelial cells by Tiam1-Rac signaling. Science 278: 1464–1466.

    Article  CAS  PubMed  Google Scholar 

  • Itoh RE, Kurokawa K, Ohba Y, Yoshizaki H, Mochizuki N, Matsuda M . (2002). Activation of rac and cdc42 video imaged by fluorescent resonance energy transfer-based single-molecule probes in the membrane of living cells. Mol Cell Biol 22: 6582–6591.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Iyer VR, Eisen MB, Ross DT, Schuler G, Moore T, Lee JC et al. (1999). The transcriptional program in the response of human fibroblasts to serum. Science 283: 83–87.

    Article  CAS  PubMed  Google Scholar 

  • Keely PJ, Westwick JK, Whitehead IP, Der CJ, Parise LV . (1997). Cdc42 and Rac1 induce integrin-mediated cell motility and invasiveness through PI(3)K. Nature 390: 632–636.

    Article  CAS  PubMed  Google Scholar 

  • Kurokawa K, Itoh RE, Yoshizaki H, Nakamura YO, Matsuda M . (2004). Coactivation of Rac1 and Cdc42 at lamellipodia and membrane ruffles induced by epidermal growth factor. Mol Biol Cell 15: 1003–1010.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ma PC, Maulik G, Christensen J, Salgia R . (2003). c-Met: structure, functions and potential for therapeutic inhibition. Cancer Metastasis Rev 22: 309–325.

    Article  CAS  PubMed  Google Scholar 

  • Matsuda M, Kobayashi Y, Masuda S, Adachi M, Watanabe T, Yamashita JK et al. (2008). Identification of adherens junction-associated GTPase activating proteins by the fluorescence localization-based expression cloning. Exp Cell Res 314: 939–949.

    Article  CAS  PubMed  Google Scholar 

  • Medico E, Gentile A, Lo CC, Williams TA, Gambarotta G, Trusolino L et al. (2001). Osteopontin is an autocrine mediator of hepatocyte growth factor-induced invasive growth. Cancer Res 61: 5861–5868.

    CAS  PubMed  Google Scholar 

  • Medico E, Mongiovi AM, Huff J, Jelinek MA, Follenzi A, Gaudino G et al. (1996). The tyrosine kinase receptors Ron and Sea control ‘scattering’ and morphogenesis of liver progenitor cells in vitro. Mol Biol Cell 7: 495–504.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Moon SY, Zheng Y . (2003). Rho GTPase-activating proteins in cell regulation. Trends Cell Biol 13: 13–22.

    Article  CAS  PubMed  Google Scholar 

  • Naldini L, Vigna E, Bardelli A, Follenzi A, Galimi F, Comoglio PM . (1995). Biological activation of pro-HGF (hepatocyte growth factor) by urokinase is controlled by a stoichiometric reaction. J Biol Chem 270: 603–611.

    Article  CAS  PubMed  Google Scholar 

  • Pankov R, Endo Y, Even-Ram S, Araki M, Clark K, Cukierman E et al. (2005). A Rac switch regulates random versus directionally persistent cell migration. J Cell Biol 170: 793–802.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Parker PJ . (1995). Intracellular signalling. PI 3-kinase puts GTP on the Rac. Curr Biol 5: 577–579.

    Article  CAS  PubMed  Google Scholar 

  • Peck J, Douglas G, Wu CH, Burbelo PD . (2002). Human RhoGAP domain-containing proteins: structure, function and evolutionary relationships. FEBS Lett 528: 27–34.

    Article  CAS  PubMed  Google Scholar 

  • Ponzetto C, Bardelli A, Zhen Z, Maina F, dalla ZP, Giordano S et al. (1994). A multifunctional docking site mediates signaling and transformation by the hepatocyte growth factor/scatter factor receptor family. Cell 77: 261–271.

    Article  CAS  PubMed  Google Scholar 

  • Ponzetto C, Zhen Z, Audero E, Maina F, Bardelli A, Basile ML et al. (1996). Specific uncoupling of GRB2 from the Met receptor. Differential effects on transformation and motility. J Biol Chem 271: 14119–14123.

    Article  CAS  PubMed  Google Scholar 

  • Ridley AJ, Comoglio PM, Hall A . (1995). Regulation of scatter factor/hepatocyte growth factor responses by Ras, Rac, and Rho in MDCK cells. Mol Cell Biol 15: 1110–1122.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ridley AJ, Self AJ, Kasmi F, Paterson HF, Hall A, Marshall CJ et al. (1993). rho family GTPase activating proteins p190, bcr and rhoGAP show distinct specificities in vitro and in vivo. EMBO J 12: 5151–5160.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rosen EM, Meromsky L, Goldberg I, Bhargava M, Setter E . (1990). Studies on the mechanism of scatter factor. Effects of agents that modulate intracellular signal transduction, macromolecule synthesis and cytoskeleton assembly. J Cell Sci 96 (Part 4): 639–649.

    CAS  PubMed  Google Scholar 

  • Royal I, Lamarche-Vane N, Lamorte L, Kaibuchi K, Park M . (2000). Activation of cdc42, rac, PAK, and rho-kinase in response to hepatocyte growth factor differentially regulates epithelial cell colony spreading and dissociation. Mol Biol Cell 11: 1709–1725.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Royal I, Park M . (1995). Hepatocyte growth factor-induced scatter of Madin-Darby canine kidney cells requires phosphatidylinositol 3-kinase. J Biol Chem 270: 27780–27787.

    Article  CAS  PubMed  Google Scholar 

  • Sander EE, ten Klooster JP, van DS, van der Kammen RA, Collard JG . (1999). Rac downregulates Rho activity: reciprocal balance between both GTPases determines cellular morphology and migratory behavior. J Cell Biol 147: 1009–1022.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sander EE, van DS, ten Klooster JP, Reid T, van der Kammen RA, Michiels F et al. (1998). Matrix-dependent Tiam1/Rac signaling in epithelial cells promotes either cell–cell adhesion or cell migration and is regulated by phosphatidylinositol 3-kinase. J Cell Biol 143: 1385–1398.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Seoh ML, Ng CH, Yong J, Lim L, Leung T . (2003). ArhGAP15, a novel human RacGAP protein with GTPase binding property. FEBS Lett 539: 131–137.

    Article  CAS  PubMed  Google Scholar 

  • Takenawa T, Miki H . (2001). WASP and WAVE family proteins: key molecules for rapid rearrangement of cortical actin filaments and cell movement. J Cell Sci 114: 1801–1809.

    CAS  PubMed  Google Scholar 

  • Trusolino L, Bertotti A, Comoglio PM . (2001). A signaling adapter function for alpha6beta4 integrin in the control of HGF-dependent invasive growth. Cell 107: 643–654.

    Article  CAS  PubMed  Google Scholar 

  • Trusolino L, Comoglio PM . (2002). Scatter-factor and semaphorin receptors: cell signalling for invasive growth. Nat Rev Cancer 2: 289–300.

    Article  CAS  PubMed  Google Scholar 

  • Wheeler AP, Wells CM, Smith SD, Vega FM, Henderson RB, Tybulewicz VL et al. (2006). Rac1 and Rac2 regulate macrophage morphology but are not essential for migration. J Cell Sci 119: 2749–2757.

    Article  CAS  PubMed  Google Scholar 

  • Zandy NL, Playford M, Pendergast AM . (2007). Abl tyrosine kinases regulate cell–cell adhesion through Rho GTPases. Proc Natl Acad Sci USA 104: 17686–17691.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zeng Q, Chen S, You Z, Yang F, Carey TE, Saims D et al. (2002). Hepatocyte growth factor inhibits anoikis in head and neck squamous cell carcinoma cells by activation of ERK and Akt signaling independent of NFkappa B. J Biol Chem 277: 25203–25208.

    Article  CAS  PubMed  Google Scholar 

  • Zhang Z, Wu C, Wang S, Huang W, Zhou Z, Ying K et al. (2002). Cloning and characterization of ARHGAP12, a novel human rhoGAP gene. Int J Biochem Cell Biol 34: 325–331.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Antonella Cignetto and Michela Bruno for secretarial assistance; Daniela Cantarella, Raffaella Albano, Laura Palmas and Solange Tienga for technical assistance and Giorgio Scita, Livio Trusolino and Lucia Biondi for helpful discussion and feedback. A particular thank to Luigi Naldini and Michele De Palma for early feedback on the design of viral traps. AG and AB are recipients of a fellowship from FIRC. This work was supported by grants from AIRC, EC (TRANSFOG EC Integrated project contract no. 503438), FIRB, MIUR, Ricerca Finalizzata, Regione Piemonte and San Paolo to EM and PMC.

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  1. L D'Alessandro

    Present address: 4Current address: German Cancer Research Center, Systems Biology of Signal Transduction, Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany,

Authors and Affiliations

  1. Laboratory of Functional Genomics, The Oncogenomics Center, Institute for Cancer Research and Treatment, University of Turin Medical School, Turin, Italy

    A Gentile, L D'Alessandro, L Lazzari, B Martinoglio, A Mira & E Medico

  2. Division of Molecular Oncology, Institute for Cancer Research and Treatment, University of Turin Medical School, Turin, Italy

    A Bertotti & P M Comoglio

  3. Division of Molecular Angiogenesis, Institute for Cancer Research and Treatment, University of Turin Medical School, Turin, Italy

    L Lanzetti

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Gentile, A., D'Alessandro, L., Lazzari, L. et al. Met-driven invasive growth involves transcriptional regulation of Arhgap12. Oncogene 27, 5590–5598 (2008). https://doi.org/10.1038/onc.2008.173

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