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ISSN 1007-7626 CN 11-3870 / Q http / / cjbmb bjmu edu cn Chinese Journal of Biochemistry and Molecular Biology 2011 4 27 4 300 ~ 304 Gab2 * 310058 Gab2 Gabs Gab2 SH2 SHP2 / Ras / ERK PI3K / AKT GAB2 ErbB2 / Neu Gab2 Gab2 Q291 Gab2 Signaling in Breast Cancer ZHANG Shan-Shan KE Yue-Hai * School of Medicine Zhejiang University Hangzhou 310058 China Abstract Gab2 is a key member of growth factor receptor-bound protein 2 Grb2 -associated binder Gab family proteins which transduces cellular signals between receptors and intracellular downstream effectors and provide platforms for protein interactions As a scaffolding adaptor Gab2 is usually activated by protein-tyrosine kinases PTKs integrating signals from a wide variety of extracellular stimuli When phosphorylated by PTKs Gab2 recruits several SH2 domain-containing proteins and subsequently triggers various downstream signal events such as SHP2 / Ras / ERK and PI3K / AKT playing an important role in cell growth differentiation apoptosis and migration Recently it has been reported that Gab2 is overexpressed in some primary mammary tumors and human breast cancer cell lines Gab2 mediated molecular events have been closely associated with mammary tumorigenesis and metastasis in coordination with oncogenic activities ErbB2 / Neu etc This review mainly focuses on the structure involvement in signal transduction of Gab2 protein and its regulation in breast cancer Key words scaffolding protein growth factor receptor-bound protein 2 Gab2 signal transduction breast cancer protein tyrosine kinases PTKs protein tyrosine phosphatases PTPs B / T 1 2010-12-06 2010-01-25 No 2009R10005 * 0571-88208713 E-mail yke@ zju edu cn Received December 6 2010 Accepted January 25 2011 Supported by Zhejiang Qianjiang Talents Project No 2009R10005 * Corresponding author 0571-88208713 E-mail yke@ zju edu cn

4 Gab2 301 SH2 Src homology-2 PTB 1 Gab2 phospho-tyrosine binding GAB2 11q14 1 97 ~ 100 kd Gab2 3 N PH PRD proline-rich 25 7 Gabs Grb2-associated binder domain C Gab family proteins Fig 1 PH Gabs Gab1 ~ Gab4 DOS daughter of sevenless Ne-Gab phosphatidyl-inositol-phosphates PIPS PIP 3 Nematostella-Gab Soc1 suppressor of clear-1 3 Gab2 phosphatidylinositol-3 4-bisphosphate PI 4 5 P 2 PH pleckstrin homology phosphatidylinositol-4 5-bisphosphate 8 SH2 SH3 Src homology-3 phosphatidylinositol-3 4 5-trisphosphate PI 3 4 P 2 Gab2 PRD PXXP Proline-rich sequences SH2 Gab2 SH3 Gabs SH3 Grb2 C 4-6 Gab2 GAB2 SH2 SHP2 SH2 Gab2 domain-containing protein-tyrosine phosphatase-2 Gab2 PI3K phosphatidyl-inositol-3-kinase p85 PLCγ phospholipase γ Crk Sarcoma virus CT10 oncogene homolog4 9 Fig 1 Structure of Gab2 scaffolding protein N-terminal PH domain is highly conversed its binding to PIPS is involved in membrane recruitment of Gab2 The central PRD contains numerous PXXP motifs mediating the interaction with SH3 domain-containing proteins such as Grb2 C-terminal multiple tyrosines are the binding sites for SH2-domain containing proteins such as SHP2 and p85 Grb2 SH2 2 Gab2 Gab2 2 Grb2 2 1 Gab2 PXXPXR Gab2 2 PX3RX2KP Grb2 2 PH SH3 Gab2 10 11 Gab2 5 6 Grb2 Grb2 Gab2 IL2 Grb2 2 SH3 1 SH2 interleukin-2 IL3 interleukin-3 Grb2 SHC Src

302 27 homology 2 domain-containing Gab2 Gab2 Grb2 SHC Gab2 12 SH2 SH3 SHP2 SH2 SHP2 2 2 Gab2 Gab2 Gab2 / SHP2 RAS / ERK SHP2 / Ras / ERK PI3K / AKT c-kit Gab2 Rac β1- PLCγ Crk GC-GAP Ras-GTPase activating protein STAT3 signal transducer and activator-3 STAT5 signal transducer and activator-5 Fig 2 Fig 2 Involvement of Gab2 protein in signal transduction 3 Characteristic of this mechanism is that the activated surface receptors serve as binding sites for SH2-domain of Grb2 which then binds to Gab2 via its SH3 domain Activated receptors lead to tyrosine phosphorylation of Gab2 protein and subsequent recruitment of SH2-domain containing effectors such as SHP2 p85 PLCγ Crk and STATs PH-domain confers recruitment of Gab2 to plasma membrane patches enriched in PIPS Gab2 3 YXXM PI3K 2 2 1 SHP2 / Ras / ERK SHP2 Gab2 2 SH2 N SH2 C Gab2 protein tyrosine phosphatase PTP SHP2 3 1 Gab2 13 Gab2 2 SHP2 Gab2 acutemyeloid leukemia AML / bisphosphoryl tyrosine-based activation motif BTAM SHP2 2 SH2 14 Ras / ERK Gab2 PI3K 2 2 2 PI3K / AKT GABs / PI3K Gab2 Gab2 p85 6 PI3K PIP S PIPS Gab2 PH Gab2 PI3K Gab2 9 B Gab1 / Gab2 SHP2 / ERK PI3K / AKT PI3K Gab2-SHP2 6 Gab2 / SHP2 / ERK PI3K 3 Gab2 Gab2 B / T Gab2 Gab2 Gab2 myelodysplastic syndrome MDS GAB2 SHP2 15

4 Gab2 303 20% ~ 30% Gab2 16 ~ 18 10% ~ 15% estrogen receptor 25 Gab2 + ER + RhoA Ras homolog gene family member A 19 26 Gab2 Gab2 20 Gab2 27 Gab2 ErbB2 / Neu 20 21 GAB2 Gab2 Gab2 Daly 19 Gab2 Gab2 Gab2 Ras / ERK PI3K / AKT ER ER + Gab2 Gab2 Sf-STK SHP2 PTP11 2 PTP Gab2 Gab2 17 epidermal growth factor receptor EGFR human epidermal growth factor receptor HER ErbB2 Neu HER2 20 Ke 24 ErbB2 / Neu Src v-sea Gab2 SHP2 SHP2 PTPN11 EGFR 28 2 SHP2 ErbB3 HER3 ErbB4 HER4 SHP2 ErbB2 / Neu Gab2 / SHP2 22 ErbB2 / Neu 4 20% ~ 30% ErbB2 / Neu Gab2 23 Gab2 ErbB2 / Neu GAB2 ErbB2 / Neu Gabs Gab2 Gabs ErbB2 / Neu Gab2 Gab2 ErbB2 / Neu 21 24 Gab2 3 2 Gab2 ErbB2 / Neu Gab2 Gab2 MCF10A RAS-ERK PI3K-AKT Gab2

304 27 GAB2 SHP2 J J Biol Chem 2001 Gab2 References 1 Ostman A Hellberg C Bohmer F D Protein-tyrosine phosphatases and cancer J Nat Rev Cancer 2006 6 4 307-320 2 Eulenfeld R Schaper F A new mechanism for the regulation of Gab1 recruitment to the plasma membrane J J Cell Sci 2009 122 Pt 1 55-64 3 Wohrle F U Daly R J Brummer T Function regulation and pathological roles of the Gab / DOS docking proteins J Cell Commun Signal 2009 7 22 4 Nishida K Hirano T The role of Gab family scaffolding adapter proteins in the signal transduction of cytokine and growth factor receptors J Cancer Sci 2003 94 12 1029-1033 5 Gu H Neel B G The Gab in signal transduction J Trends Cell Biol 2003 13 3 122-130 6 Maus M Medgyesi D Kovesdi D et al Grb2 associated binder 2 couples B-cell receptor to cell survival J Cell Signal 2009 21 2 220-227 7 Pan X L Ren R J Wang G et al The Gab2 in signal transduction and its potential role in the pathogenesis of Alzheimer s disease J Neurosci Bull 2010 26 3 241-246 8 Franke T F PI3K / Akt getting it right matters J Oncogene 2008 27 50 6473-6488 9 Zhang T T Li H Cheung S M et al Phosphoinositide 3- kinase-regulated adapters in lymphocyte activation J Immunol Rev 2009 232 1 255-272 10 Lock L S Royal I Naujokas M A et al Identification of an atypical Grb2 carboxyl-terminal SH3 domain binding site in Gab docking proteins reveals Grb2-dependent and-independent recruitment of Gab1 to receptor tyrosine kinases J J Biol Chem 2000 275 40 31536-31545 11 Harkiolaki M Tsirka T Lewitzky M et al Distinct binding modes of two epitopes in Gab2 that interact with the SH3C domain of Grb2 J Structure 2009 17 6 809-822 12 Gu H Maeda H Moon J J et al New role for Shc in activation of the phosphatidylinositol 3-kinase / Akt pathway J Mol Cell Biol 2000 20 19 7109-7120 13 Neel B G Gu H Pao L The Shp ing news SH2 domaincontaining tyrosine phosphatases in cell signaling J Trends Biochem Sci 2003 28 6 284-293 14 Cunnick J M Mei L Doupnik C A et al Phosphotyrosines 627 and 659 of Gab1 constitute a bisphosphoryl tyrosine-based activation motif BTAM conferring binding and activation of 276 26 24380-24387 15 Szepetowski P Ollendorff V Grosgeorge J et al DNA amplification at 11q13 5-q14 in human breast cancer J Oncogene 1992 7 12 2513-2517 16 Brown L A Kalloger S E Miller M A et al Amplification of 11q13 in ovarian carcinoma J Genes Chromosomes Cancer 2008 47 6 481-489 17 Bocanegra M Bergamaschi A Kim Y H et al Focal amplification and oncogene dependency of GAB2 in breast cancer J Oncogene 2010 29 5 774-779 18 Bekri S Adelaide J Merscher S et al Detailed map of a region commonly amplified at 11q13--> q14 in human breast carcinoma J Cytogenet Cell Genet 1997 79 1-2 125-131 19 Daly R J Gu H Parmar J et al The docking protein Gab2 is overexpressed and estrogen regulated in human breast cancer J Oncogene 2002 21 33 5175-5181 20 Fleuren E D O Toole S Millar E K et al Overexpression of the oncogenic signal transducer Gab2 occurs early in breast cancer development J Int J Cancer 2010 127 6 1486-1492 21 Bentires-Alj M Gil S G Chan R et al A role for the scaffolding adapter GAB2 in breast cancer J Nat Med 2006 12 1 114-121 22 Citri A Yarden Y EGF-ERBB signalling towards the systems level J Nat Rev Mol Cell Biol 2006 7 7 505-516 23 Slamon D J Leyland-Jones B Shak S et al Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2 J N Engl J Med 2001 344 11 783-792 24 Ke Y Wu D Princen F et al Role of Gab2 in mammary tumorigenesis and metastasis J Oncogene 2007 26 34 4951-4960 25 Mira A Isella C Renzulli T et al The GAB2 signaling scaffold promotes anchorage independence and drives a transcriptional response associated with metastatic progression of breast cancer J Oncogene 2009 28 50 4444-4455 26 Abreu M T Hughes W E Mele K et al Gab2 regulates cytoskeletal organization and migration of mammary epithelial cells by modulating RhoA activation J Mol Biol Cell 2011 22 1 105-116 27 Frigault M M Naujokas M A Park M Gab2 requires membrane targeting and the Met binding motif to promote lamellipodia cell scatter and epithelial morphogenesis downstream from the Met receptor J J Cell Physiol 2008 214 3 694-705 28 Zhou X Coad J Ducatman B et al SHP2 is up-regulated in breast cancer cells and in infiltrating ductal carcinoma of the breast implying its involvement in breast oncogenesis J Histopathology 2008 53 4 389-402