Abstract
E-cadherin and its cytoplasmic partners, catenins, mediate epithelial cell–cell adhesion. Disruption of this adhesion allows cancer cells to invade and metastasize. Aberrant activation of the Src tyrosine kinase disrupts cell–cell contacts through an E-cadherin/catenin-dependent mechanism. Previously we showed that Rack1 regulates the growth of colon cells by suppressing Src activity at G1 and mitotic checkpoints, and in the intrinsic apoptotic and Akt cell survival pathways. Here we show that Rack1, partly by inhibiting Src, promotes cell–cell adhesion and reduces the invasive potential of colon cancer cells. Rack1 stabilizes E-cadherin and catenins at cell–cell contacts by inhibiting the Src phosphorylation of E-cadherin, the ubiquitination of E-cadherin by the E3 ligase Hakai and the endocytosis of E-cadherin. Upon depletion and restoration of extracellular calcium, Rack1 facilitates the re-assembly of E-cadherin-containing cell–cell contacts. Rack1 also blocks HGF-induced endocytosis of E-cadherin, disruption of cell–cell contacts and cell scatter. Our results uncover a novel function of Rack1 in maintaining the junctional homeostasis of intestinal epithelial cells by regulation of the Src- and growth factor-induced endocytosis of E-cadherin.
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References
Avizienyte E, Fincham VJ, Brunton VG, Frame MC . (2004). Src SH3/2 domain-mediated peripheral accumulation of Src and phospho-myosin is linked to deregulation of E-cadherin and the epithelial–mesenchymal transition. Mol Biol Cell 15: 2794–2803.
Avizienyte E, Frame MC . (2005). Src and FAK signalling controls adhesion fate and the epithelial-to-mesenchymal transition. Curr Opin Cell Biol 17: 542–547.
Avizienyte E, Wyke AW, Jones RJ, McLean GW, Westhoff MA, Brunton VG et al. (2002). Src-induced de-regulation of E-cadherin in colon cancer cells requires integrin signalling. Nat Cell Biol 4: 632–638.
Behrens J, Vakaet L, Friis R, Winterhager E, Van Roy F, Mareel MM et al. (1993). Loss of epithelial differentiation and gain of invasiveness correlates with tyrosine phosphorylation of the E-cadherin/beta-catenin complex in cells transformed with a temperature-sensitive v-SRC gene. J Cell Biol 120: 757–766.
Bertotti A, Bracco C, Girolami F, Torti D, Gastaldi S, Galimi F et al. (2010). Inhibition of Src impairs the growth of met-addicted gastric tumors. Clin Cancer Res 16: 3933–3943.
Berx G, van Roy F . (2009). Involvement of members of the cadherin superfamily in cancer. Cold Spring Harb Perspect Biol 1: a003129.
Brady-Kalnay SM, Mourton T, Nixon JP, Pietz GE, Kinch M, Chen H et al. (1998). Dynamic interaction of PTPmu with multiple cadherins in vivo. J Cell Biol 141: 287–296.
Brady-Kalnay SM, Rimm DL, Tonks NK . (1995). Receptor protein tyrosine phosphatase PTPmu associates with cadherins and catenins in vivo. J Cell Biol 130: 977–986.
Bryant DM, Stow JL . (2004). The ins and outs of E-cadherin trafficking. Trends Cell Biol 14: 427–434.
Chang BY, Chiang M, Cartwright CA . (2001). The interaction of Src and RACK1 is enhanced by activation of protein kinase C and tyrosine phosphorylation of RACK1. J Biol Chem 276: 20346–20356.
Chang BY, Conroy KB, Machleder EM, Cartwright CA . (1998). RACK1, a receptor for activated C kinase and a homolog of the beta subunit of G proteins, inhibits activity of src tyrosine kinases and growth of NIH 3T3 cells. Mol Cell Biol 18: 3245–3256.
Chang BY, Harte RA, Cartwright CA . (2002). RACK1: a novel substrate for the Src protein-tyrosine kinase. Oncogene 21: 7619–7629.
Chattopadhyay N, Wang Z, Ashman LK, Brady-Kalnay SM, Kreidberg JA . (2003). alpha3beta1 Integrin–CD151, a component of the cadherin–catenin complex, regulates PTPmu expression and cell–cell adhesion. J Cell Biol 163: 1351–1362.
Cox EA, Bennin D, Doan AT, O'Toole T, Huttenlocher A . (2003). RACK1 regulates integrin-mediated adhesion, protrusion, and chemotactic cell migration via its Src-binding site. Mol Biol Cell 14: 658–669.
Delva E, Kowalczyk AP . (2009). Regulation of cadherin trafficking. Traffic 10: 259–267.
Doan AT, Huttenlocher A . (2007). RACK1 regulates Src activity and modulates paxillin dynamics during cell migration. Exp Cell Res 313: 2667–2679.
Emaduddin M, Bicknell DC, Bodmer WF, Feller SM . (2008). Cell growth, global phosphotyrosine elevation, and c-Met phosphorylation through Src family kinases in colorectal cancer cells. Proc Natl Acad Sci USA 105: 2358–2362.
Fan S, Meng Q, Laterra JJ, Rosen EM . (2009). Role of Src signal transduction pathways in scatter factor-mediated cellular protection. J Biol Chem 284: 7561–7577.
Fomenkov A, Zangen R, Huang YP, Osada M, Guo Z, Fomenkov T et al. (2004). RACK1 and stratifin target DeltaNp63alpha for a proteasome degradation in head and neck squamous cell carcinoma cells upon DNA damage. Cell Cycle 3: 1285–1295.
Frame MC . (2004). Newest findings on the oldest oncogene; how activated src does it. J Cell Sci 117: 989–998.
Fujita Y, Krause G, Scheffner M, Zechner D, Leddy HE, Behrens J et al. (2002). Hakai, a c-Cbl-like protein, ubiquitinates and induces endocytosis of the E-cadherin complex. Nat Cell Biol 4: 222–231.
Gumbiner BM . (2005). Regulation of cadherin-mediated adhesion in morphogenesis. Nat Rev Mol Cell Biol 6: 622–634.
Hartsock A, Nelson WJ . (2008). Adherens and tight junctions: structure, function and connections to the actin cytoskeleton. Biochim Biophys Acta 1778: 660–669.
Hellberg CB, Burden-Gulley SM, Pietz GE, Brady-Kalnay SM . (2002). Expression of the receptor protein-tyrosine phosphatase, PTPmu, restores E-cadherin-dependent adhesion in human prostate carcinoma cells. J Biol Chem 277: 11165–11173.
Herynk MH, Zhang J, Parikh NU, Gallick GE . (2007). Activation of Src by c-Met overexpression mediates metastatic properties of colorectal carcinoma cells. J Exp Ther Oncol 6: 205–217.
Irby RB, Yeatman TJ . (2002). Increased Src activity disrupts cadherin/catenin-mediated homotypic adhesion in human colon cancer and transformed rodent cells. Cancer Res 62: 2669–2674.
Janda E, Nevolo M, Lehmann K, Downward J, Beug H, Grieco M . (2006). Raf plus TGFbeta-dependent EMT is initiated by endocytosis and lysosomal degradation of E-cadherin. Oncogene 25: 7117–7130.
Jeanes A, Gottardi CJ, Yap AS . (2008). Cadherins and cancer: how does cadherin dysfunction promote tumor progression? Oncogene 27: 6920–6929.
Kiely PA, Leahy M, O'Gorman D, O'Connor R . (2005). RACK1-mediated integration of adhesion and insulin-like growth factor I (IGF-I) signaling and cell migration are defective in cells expressing an IGF-I receptor mutated at tyrosines 1250 and 1251. J Biol Chem 280: 7624–7633.
Kim JB, Islam S, Kim YJ, Prudoff RS, Sass KM, Wheelock MJ et al. (2000). N-cadherin extracellular repeat 4 mediates epithelial to mesenchymal transition and increased motility. J Cell Biol 151: 1193–1206.
Kowalczyk AP, Reynolds AB . (2004). Protecting your tail: regulation of cadherin degradation by p120-catenin. Curr Opin Cell Biol 16: 522–527.
Lai AZ, Abella JV, Park M . (2009). Crosstalk in Met receptor oncogenesis. Trends Cell Biol 19: 542–551.
Leroy C, Fialin C, Sirvent A, Simon V, Urbach S, Poncet J et al. (2009). Quantitative phosphoproteomics reveals a cluster of tyrosine kinases that mediates SRC invasive activity in advanced colon carcinoma cells. Cancer Res 69: 2279–2286.
Li Y, Peart MJ, Prives C . (2009). Stxbp4 regulates DeltaNp63 stability by suppression of RACK1-dependent degradation. Mol Cell Biol 29: 3953–3963.
Lipsich LA, Lewis AJ, Brugge JS . (1983). Isolation of monoclonal antibodies that recognize the transforming proteins of avian sarcoma viruses. J Virol 48: 352–360.
Liu YV, Baek JH, Zhang H, Diez R, Cole RN, Semenza GL . (2007). RACK1 competes with HSP90 for binding to HIF-1alpha and is required for O(2)-independent and HSP90 inhibitor-induced degradation of HIF-1alpha. Mol Cell 25: 207–217.
Lu Z, Ghosh S, Wang Z, Hunter T . (2003). Downregulation of caveolin-1 function by EGF leads to the loss of E-cadherin, increased transcriptional activity of beta-catenin, and enhanced tumor cell invasion. Cancer Cell 4: 499–515.
Mamidipudi V, Cartwright CA . (2009). A novel pro-apoptotic function of RACK1: suppression of Src activity in the intrinsic and Akt pathways. Oncogene 28: 4421–4433.
Mamidipudi V, Dhillon NK, Parman T, Miller LD, Lee KC, Cartwright CA . (2007). RACK1 inhibits colonic cell growth by regulating Src activity at cell cycle checkpoints. Oncogene 26: 2914–2924.
Mamidipudi V, Zhang J, Lee KC, Cartwright CA . (2004). RACK1 regulates G1/S progression by suppressing Src kinase activity. Mol Cell Biol 24: 6788–6798.
McCahill A, Warwicker J, Bolger GB, Houslay MD, Yarwood SJ . (2002). The RACK1 scaffold protein: a dynamic cog in cell response mechanisms. Mol Pharmacol 62: 1261–1273.
Mosesson Y, Mills GB, Yarden Y . (2008). Derailed endocytosis: an emerging feature of cancer. Nat Rev Cancer 8: 835–850.
Mourton T, Hellberg CB, Burden-Gulley SM, Hinman J, Rhee A, Brady-Kalnay SM . (2001). The PTPmu protein-tyrosine phosphatase binds and recruits the scaffolding protein RACK1 to cell–cell contacts. J Biol Chem 276: 14896–14901.
Ohgaki R, Fukura N, Matsushita M, Mitsui K, Kanazawa H . (2008). Cell surface levels of organellar Na+/H+ exchanger isoform 6 are regulated by interaction with RACK1. J Biol Chem 283: 4417–4429.
Orlichenko L, Weller SG, Cao H, Krueger EW, Awoniyi M, Beznoussenko G et al. (2009). Caveolae mediate growth factor-induced disassembly of adherens junctions to support tumor cell dissociation. Mol Biol Cell 20: 4140–4152.
Owens DW, McLean GW, Wyke AW, Paraskeva C, Parkinson EK, Frame MC et al. (2000). The catalytic activity of the Src family kinases is required to disrupt cadherin-dependent cell–cell contacts. Mol Biol Cell 11: 51–64.
Palacios F, Tushir JS, Fujita Y, D'Souza-Schorey C . (2005). Lysosomal targeting of E-cadherin: a unique mechanism for the downregulation of cell–cell adhesion during epithelial to mesenchymal transitions. Mol Cell Biol 25: 389–402.
Parent A, Laroche G, Hamelin E, Parent JL . (2008). RACK1 regulates the cell surface expression of the G protein-coupled receptor for thromboxane A(2). Traffic 9: 394–407.
Rengifo-Cam W, Konishi A, Morishita N, Matsuoka H, Yamori T, Nada S et al. (2004). Csk defines the ability of integrin-mediated cell adhesion and migration in human colon cancer cells: implication for a potential role in cancer metastasis. Oncogene 23: 289–297.
Saitoh M, Shirakihara T, Miyazono K . (2009). Regulation of the stability of cell surface E-cadherin by the proteasome. Biochem Biophys Res Commun 381: 560–565.
Schechtman D, Mochly-Rosen D . (2001). Adaptor proteins in protein kinase C-mediated signal transduction. Oncogene 20: 6339–6347.
Schill NJ, Anderson RA . (2009). Out, in and back again: PtdIns(4,5)P(2) regulates cadherin trafficking in epithelial morphogenesis. Biochem J 418: 247–260.
Serrels B, Sandilands E, Serrels A, Baillie G, Houslay MD, Brunton VG et al. (2010). A complex between FAK, RACK1, and PDE4D5 controls spreading initiation and cancer cell polarity. Curr Biol 20: 1086–1092.
Shen Y, Hirsch DS, Sasiela CA, Wu WJ . (2008). Cdc42 regulates E-cadherin ubiquitination and degradation through an epidermal growth factor receptor to Src-mediated pathway. J Biol Chem 283: 5127–5137.
Skoudy A, Llosas MD, Garcia de Herreros A . (1996). Intestinal HT-29 cells with dysfunction of E-cadherin show increased pp60src activity and tyrosine phosphorylation of p120-catenin. Biochem J 317 (Part 1): 279–284.
Varghese B, Barriere H, Carbone CJ, Banerjee A, Swaminathan G, Plotnikov A et al. (2008). Polyubiquitination of prolactin receptor stimulates its internalization, postinternalization sorting, and degradation via the lysosomal pathway. Mol Cell Biol 28: 5275–5287.
Vomastek T, Iwanicki MP, Schaeffer HJ, Tarcsafalvi A, Parsons JT, Weber MJ . (2007). RACK1 targets the extracellular signal-regulated kinase/mitogen-activated protein kinase pathway to link integrin engagement with focal adhesion disassembly and cell motility. Mol Cell Biol 27: 8296–8305.
Wadhawan A, Smith C, Nicholson RI, Barrett-Lee P, Hiscox S . (2010). Src-mediated regulation of homotypic cell adhesion: implications for cancer progression and opportunities for therapeutic intervention. Cancer Treat Rev 37: 234–241.
Woodcock SA, Rooney C, Liontos M, Connolly Y, Zoumpourlis V, Whetton AD et al. (2009). SRC-induced disassembly of adherens junctions requires localized phosphorylation and degradation of the rac activator tiam1. Mol Cell 33: 639–653.
Yap AS, Crampton MS, Hardin J . (2007). Making and breaking contacts: the cellular biology of cadherin regulation. Curr Opin Cell Biol 19: 508–514.
Yeatman TJ . (2004). A renaissance for SRC. Nat Rev Cancer 4: 470–480.
Acknowledgements
We thank Toufan Parman for performing initial preliminary experiments. We are grateful to Jenny Cheng, James Nelson, Anson Lowe and Manuel Amieva for helpful discussions and reagents. We thank James Nelson for the gift of MDCK cells and the members of the Lipsick laboratory for help with confocal imaging. We thank Jenny Cheng for critical review of the data and the manuscript. This work was supported by the NIH Grant DK43743 (CAC) and the NIH Digestive Disease Center Grant DK56339.
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Swaminathan, G., Cartwright, C. Rack1 promotes epithelial cell–cell adhesion by regulating E-cadherin endocytosis. Oncogene 31, 376–389 (2012). https://doi.org/10.1038/onc.2011.242
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DOI: https://doi.org/10.1038/onc.2011.242
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