This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kapoor, G. S. Articles by O'Rourke, D. M. Search for Related Content PubMed PubMed Citation Articles by Kapoor, G. S. Articles by O'Rourke, D. M.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, January 2004, p. 823-836, Vol. 24, No. 2
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.2.823-836.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Distinct Domains in the SHP-2 Phosphatase Differentially Regulate Epidermal Growth Factor Receptor/NF-B Activation through Gab1 in Glioblastoma Cells

Gurpreet S. Kapoor,¹ Yi Zhan,¹ Gibbes R. Johnson,² and Donald M. O'Rourke^1,3*

Departments of Neurosurgery,¹ Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104,³ Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892²

Received 21 July 2003/ Returned for modification 11 September 2003/ Accepted 15 October 2003

The transcription factor nuclear factor B (NF-B) plays an important role in inflammation and cancer, is activated by a variety of stimuli including tumor necrosis factor alpha, interleukin-1, UV irradiation, and viruses, as well as receptor tyrosine kinases, such as epidermal growth factor receptor (EGFR). Although previous studies suggest that EGFR can induce NF-B, the mechanism of this activation remains unknown. In this study, we identify the components of the EGFR-induced signalosome in human glioblastoma cells required to regulate NF-B activation. Immunoprecipitation analyses with ErbB-modulated cells indicate that association between SHP-2 and Grb2-associated binder 1 (Gab1) is the critical step in the formation of the signalosome linking EGFR to NF-B activation. We also show that EGFR-induced NF-B activation is mediated by the PI3-kinase/Akt activation loop. Overexpression of SHP-2, Gab1, and myristoylated Akt significantly upregulated NF-B transcriptional activity and DNA binding activity in glioblastoma cells. Interestingly, overexpression of either one of the two SH2 domain mutants of SHP-2, R32E or R138E, slightly reduced NF-B activity relative to that of wild-type SHP-2, indicating that the SH2 domains of SHP-2 are required for EGFR-induced NF-B activation. On the other hand, ectopic overexpression of either a Gab1 mutant incapable of binding to SHP-2 (Y627F) or a phosphatase-inactive SHP-2 mutant (C459S) caused a significant increase in NF-B activity. Moreover, SHP-2 C459S-expressing cells displayed higher Gab1 phosphotyrosine content, suggesting that SHP-2 regulates Gab1 phosphorylation through its phosphatase domain, which confers a negative regulatory effect on NF-B activity. These results indicate that SHP-2/Gab1 association is critical for linking EGFR to NF-B transcriptional activity via the PI3-kinase/Akt signaling axis in glioblastoma cells and that SHP-2 acts as a dual regulator of NF-B activation.

---

* Corresponding author. Mailing address: Department of Neurosurgery, University of Pennsylvania School of Medicine, 502 Stemmler Hall, 36th and Hamilton Walk, Philadelphia, PA 19104. Phone: (215) 898-2871. Fax: (215) 898-9217. E-mail: orourked{at}mail.med.upenn.edu.

---

Molecular and Cellular Biology, January 2004, p. 823-836, Vol. 24, No. 2
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.2.823-836.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Tabet, F., Schiffrin, E. L., Callera, G. E., He, Y., Yao, G., Ostman, A., Kappert, K., Tonks, N. K., Touyz, R. M. (2008). Redox-Sensitive Signaling by Angiotensin II Involves Oxidative Inactivation and Blunted Phosphorylation of Protein Tyrosine Phosphatase SHP-2 in Vascular Smooth Muscle Cells From SHR. Circ. Res. 103: 149-158 [Abstract] [Full Text]  
• Lee, H., Kong, S. W., Park, P. J. (2008). Integrative analysis reveals the direct and indirect interactions between DNA copy number aberrations and gene expression changes. Bioinformatics 24: 889-896 [Abstract] [Full Text]  
• Kong, X.-N., Yan, H.-X., Chen, L., Dong, L.-W., Yang, W., Liu, Q., Yu, L.-X., Huang, D.-D., Liu, S.-Q., Liu, H., Wu, M.-C., Wang, H.-Y. (2007). LPS-induced down-regulation of signal regulatory protein {alpha} contributes to innate immune activation in macrophages. JEM 204: 2719-2731 [Abstract] [Full Text]  
• Lemarie, C. A., Tharaux, P.-L., Esposito, B., Tedgui, A., Lehoux, S. (2006). Transforming Growth Factor-{alpha} Mediates Nuclear Factor {kappa}B Activation in Strained Arteries. Circ. Res. 99: 434-441 [Abstract] [Full Text]  
• Sithanandam, G., Smith, G. T., Fields, J. R., Fornwald, L. W., Anderson, L. M. (2005). Alternate Paths from Epidermal Growth Factor Receptor to Akt in Malignant Versus Nontransformed Lung Epithelial Cells: ErbB3 Versus Gab1. Am. J. Respir. Cell Mol. Bio. 33: 490-499 [Abstract] [Full Text]  
• Zhan, Y., O'Rourke, D. M. (2004). SHP-2-Dependent Mitogen-Activated Protein Kinase Activation Regulates EGFRvIII but not Wild-Type Epidermal Growth Factor Receptor Phosphorylation and Glioblastoma Cell Survival. Cancer Res. 64: 8292-8298 [Abstract] [Full Text]  
• Kapoor, G. S., Kapitonov, D., O'Rourke, D. M. (2004). Transcriptional Regulation of Signal Regulatory Protein {alpha}1 Inhibitory Receptors by Epidermal Growth Factor Receptor Signaling. Cancer Res. 64: 6444-6452 [Abstract] [Full Text]