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The interplay between Eps8 and IRSp53 contributes to Src-mediated transformation

Oncogene volume 29, pages 3977–3989 (2010)Cite this article

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Abstract

As an oncoprotein, Eps8 participates in v-Src-induced cellular transformation. To delineate the underlying mechanism, we conducted a yeast two-hybrid screening and identified IRSp53S, a protein critical in cell mobilization, as one of the Eps8-binding partners from a human brain cDNA library. The association was mediated by the multiple proline-rich regions of Eps8 and the C-terminal SH3-WWB containing domains of IRSp53S. In this study, we observed that Eps8 modulated the expression of IRSp53 in v-Src-transformed cells (IV5), raising the question of whether Eps8/IRSp53 interaction was crucial in carcinogenesis. To address this issue, we generated IV5-expressing irsp53 siRNA cells. Attenuation of IRSp53 reduced cell proliferation of IV5 in culture dish and tumor formation in mice, which could be partly rescued by ectopically expressed human IRSp53S. In addition, IRSp53 knockdown impaired activity of phosphatidylinositol 3-kinase (as reflected by Pi-Ser473 AKT) and Stat3 (as reflected by Pi-Tyr705 Stat3), and reduced cyclin D1 expression that culminated to impede G1-phase cell-cycle progression. Ectopically expressed human IRSp53S, but not its Eps8-binding defective mutants (that is, Δ363 and PPPDA), rescued these defects and partly restored cell proliferation. Remarkably, through activation of Src, EGF increased the formation of Eps8/IRSp53 complex and Stat3 activation in HeLa cells. With these results, we show for the first time that IRSp53, through its interaction with Eps8, not only affects cell migration but also dictates cellular growth in cancer cells.

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References

  • Abbott MA, Wells DG, Fallon JR . (1999). The insulin receptor tyrosine kinase substrate p58/53 and the insulin receptor are components of CNS synapses. J Neurosci 19: 7300–7308.

    Article  CAS  Google Scholar 

  • Aitio O, Hellman M, Kesti T, Kleino L, Samuilova O, Paakkonen K et al. (2008). Structural basis of PxxDY motif recognition in SH3 binding. J Mol Biol 382: 167–178.

    Article  CAS  Google Scholar 

  • Blake RA, Broome MA, Liu X, Wu J, Gishizky M, Sun L et al. (2000). SU6656, a selective Src family kinase inhibitor, used to probe growth factor signaling. Mol Cell Biol 20: 9018–9027.

    Article  CAS  Google Scholar 

  • Bowman T, Broome MA, Sinibaldi D, Wharton W, Pledger WJ, Sedivy JM et al. (2001). Stat3-mediated Myc expression is required for Src transformation and PDGF-induced mitogenesis. Proc Natl Acad Sci USA 98: 7319–7324.

    Article  CAS  Google Scholar 

  • Chen YJ, Shen MR, Chen YJ, Maa MC, Leu TH . (2008). Eps8 decreases chemosensitivity and affects survival of cervical cancer patients. Mol Cancer Ther 7: 1376–1385.

    Article  CAS  Google Scholar 

  • Disanza A, Carlier MF, Stradal TE, Didry D, Frittoli E, Confalonieri S et al. (2004). Eps8 controls actin-based motility by capping the barbed ends of actin filaments. Nat Cell Biol 6: 1180–1188.

    Article  CAS  Google Scholar 

  • Funato Y, Terabayashi T, Suenaga N, Seiki M, Takenawa T, Miki H . (2004). IRSp53/Eps8 complex is important for positive regulation of Rac and cancer cell motility/invasiveness. Cancer Res 64: 5237–5244.

    Article  CAS  Google Scholar 

  • Gallo R, Provenzano C, Carbone R, Di Fiore PP, Castellani L, Falcone G et al. (1997). Regulation of the tyrosine kinase substrate Eps8 expression by growth factor, v-Src and terminal differentiation. Oncogene 15: 1929–1936.

    Article  CAS  Google Scholar 

  • Harlow E, Lane D (eds). (1988). Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, NY, pp 522–523.

    Google Scholar 

  • Lee SH, Kerff F, Chereau D, Ferron F, Klug A, Dominguez R . (2007). Structural basis for actin-binding function of missing-in-metastasis. Structure 15: 145–155.

    Article  Google Scholar 

  • Leu TH, Yeh HH, Huang CC, Chuang YC, Su SL, Maa MC . (2004). Participation of p97Eps8 in Src-mediated transformation. J Biol Chem 279: 9875–9881.

    Article  CAS  Google Scholar 

  • Maa MC, Leu TH . (1998). Vanadate-dependent FAK activation is accomplished by the sustained FAK Tyr-576/577 phosphorylation. Biochem Biophy Res Commun 251: 344–349.

    Article  CAS  Google Scholar 

  • Maa MC, Lai JR, Lin RW, Leu TH . (1999). Enhancement of tyrosyl phosphorylation and protein expression of eps8 by v-Src. Biochim Biophys Acta 1450: 341–351.

    Article  CAS  Google Scholar 

  • Maa MC, Hsieh CY, Leu TH . (2001). Overexpression of p97Eps8 leads to cellular transformation: implication of pleckstrin homology domain in p97Eps8-mediated ERK activation. Oncogene 20: 106–112.

    Article  CAS  Google Scholar 

  • Maa MC, Lee JC, Chen YJ, Chen YJ, Lee YC, Wang ST et al. (2007). Eps8 facilitates cellular growth and motility of colon cancer cells by increasing the expression and activity of focal adhesion kinase. J Biol Chem 282: 19399–19409.

    Article  CAS  Google Scholar 

  • Martin GS . (2001). Timeline: the hunting of the Src. Nat Rev Mol Cell Biol 2: 467–475.

    Article  CAS  Google Scholar 

  • Mattila PK, Pykalainen A, Saarikangas J, Paavilainen VO, Vihinen H, Jokitalo E et al. (2007). Missing-in-metastasis and IRSp53 deform PIP2-rich membranes by an inverse BAR domain-like mechanism. J Cell Biol 7: 953–964.

    Article  Google Scholar 

  • Millard TH, Bompard G, Heung MY, Dafforn TR, Scott DJ, Machesky LM et al. (2005). Structural basis of filopodia formation induced by the IRSp53/MIM homology domain of human IRSp53. EMBO J 24: 240–250.

    Article  CAS  Google Scholar 

  • Miyahara A, Okamura-Oho Y, Miyahita T, Hoshika A, Yamada M . (2003). Genomic structure and alternative splicing of the insulin receptor tyrosine kinase substrate of 53-kDa protein. J Hum Genet 48: 410–414.

    Article  CAS  Google Scholar 

  • Mongiovi AM, Romaon PR, Panni S, Mendoza M, Wong WT, Musacchio A et al. (1999). A novel peptide-SH3 interaction. EMBO J 18: 5300–5309.

    Article  CAS  Google Scholar 

  • Oda K, Shiratsuchi T, Nishimori H, Inazawa J, Yoshikawa H, Taketani Y et al. (1999). Identification of BAIAP2 (BAI-associated protein 2), a novel human homologue of hamster IRSp53, whose SH3 interacts with the cytoplasmic domain of BAI1. Cytogenet Cell Genet 84: 75–82.

    Article  CAS  Google Scholar 

  • Okamura-Oho Y, Miyashita T, Ohmi K, Yamada M . (1999). Dentatorubral-pallidoluysian atrophy protein interacts through a proline-rich region near polyglutamine with the SH3 domain of an insulin receptor tyrosine kinase substrate. Hum Mol Genet 8: 947–957.

    Article  CAS  Google Scholar 

  • Okamura-Oho Y, Miyashita T, Yamada M . (2001). Distinctive tissue distribution and phosphorylation of IRSp53 isoforms. Biochem Biophys Res Commun 289: 957–960.

    Article  CAS  Google Scholar 

  • Scita G, Nordstrom J, Carbone R, Tenca P, Giardina G, Gutkind S et al. (1999). EPS8 and E3B1 transduce signals from Ras to Rac. Nature 401: 290–293.

    Article  CAS  Google Scholar 

  • Scita G, Confalonieri S, Lappalainen P, Suetsugu S . (2008). IRSp53: crossing the road of membrane and actin dynamics in the formation of membrane protrusions. Trends Cell Biol 18: 52–60.

    Article  CAS  Google Scholar 

  • Silva CM . (2004). Role of STATs as downstream signal transducers in Src family kinase-mediated tumorigensis. Oncogene 23: 8017–8023.

    Article  CAS  Google Scholar 

  • Suetsugu S, Murayama K, Sakamoto A, Hanawa-Suetsugu K, Seto A, Oikawa T et al. (2006). The RAC binding domain/IRSp53-MIM homology domain of IRSp53 induces RAC-dependent membrane deformation. J Biol Chem 46: 35347–35358.

    Article  Google Scholar 

  • Xu M, Shorts-Cary L, Knox AJ, Kleinsmidt-DeMasters B, Lillehei K, Wierman ME . (2009). Epidermal growth factor receptor pathway substrate 8 is overexpressed in human pituitary tumors: role in proliferation and survival. Endocrinology 150: 2064–2071.

    Article  CAS  Google Scholar 

  • Yap LF, Jenei V, Robinson CM, Moutasim K, Benn TM, Threadgold SP et al. (2009). Upregulation of Eps8 in oral squamous cell carcinoma promotes cell migration and invasion through integrin-dependent Rac1 activation. Oncogene 28: 2524–2534.

    Article  CAS  Google Scholar 

  • Yeh TC, Ogawa W, Danielsen AG, Roth RA . (1996). Characterization and cloning of a 58/53-kDa substrate of the insulin receptor tyrosine kinase. J Biol Chem 271: 2921–2928.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by National Science Council grants to M-CM (NSC98-2311-B-039-002-MY3) and T-HL (NSC97-2320-B-006-024-MY3). Additional support came from NHRI (NHRI-EX-98-9828BI to T-H Leu), Taiwan Department of Health Clinical Trial and Research Center of Excellence (DOH99-TD-B-111-004 to M-C Maa) and China Medical University (CMU98-C-05 to M-C Maa). The DNA constructs of pLKO.1-mirsp53-b and pLKO.1-mirsp53-c were provided by the National RNAi Core Facility located at the Institute of Molecular Biology/Genomic Research Center, Academia Sinica, supported by the National Research Program for Genomic Medicine Grants of NSC (NSC 94-3112-B-001-003 and NSC 94-3112-B-001-018-Y).

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Authors and Affiliations

  1. Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    P-S Liu & T-H Leu

  2. Department of Pharmacology, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    T-H Jong & T-H Leu

  3. Institute of Medical Science, China Medical University, Taichung, Taiwan

    M-C Maa

  4. Institute of Molecular Systems Biomedicine, China Medical University, Taichung, Taiwan

    M-C Maa

  5. Center for Gene Regulation and Signal Transduction Research, National Cheng Kung University, Tainan, Taiwan

    T-H Leu

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  1. P-S Liu
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Correspondence to M-C Maa or T-H Leu.

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Liu, PS., Jong, TH., Maa, MC. et al. The interplay between Eps8 and IRSp53 contributes to Src-mediated transformation. Oncogene 29, 3977–3989 (2010). https://doi.org/10.1038/onc.2010.144

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