Perinuclear localization of the protein-tyrosine phosphatase SHP-1 and inhibition of epidermal growth factor-stimulated STAT1/3 activation in A431 cells

• Published in: European journal of cell biology Vol. 79; no. 4; pp. 261 - 271
• Main Authors: Tenev, Tencho, Böhmer, Sylvia-Annette, Kaufmann, Roland, Frese, Steffen, Bittorf, Thomas, Beckers, Thomas, Böhmer, Frank-D.
• Format: Journal Article
• Language: English
• Published: Germany Elsevier GmbH 01-04-2000
• Subjects: 3T3 Cells; Animals; Calcimycin - pharmacology; Cell Line; Cell Nucleus - metabolism; DNA, Complementary - metabolism; DNA-Binding Proteins - metabolism; Enzyme Activation; Epidermal Growth Factor - metabolism; epidermal growth factor receptor; Humans; Immunohistochemistry; Intracellular Signaling Peptides and Proteins; Ionophores - pharmacology; Keratinocytes - metabolism; localization; Mice; Protein Structure, Tertiary; Protein Synthesis Inhibitors - pharmacology; Protein tyrosine phosphatase; Protein Tyrosine Phosphatase, Non-Receptor Type 11; Protein Tyrosine Phosphatase, Non-Receptor Type 6; Protein Tyrosine Phosphatases - metabolism; Receptor, Epidermal Growth Factor - metabolism; regulatable expression system; SH2 Domain-Containing Protein Tyrosine Phosphatases; SHP-1; Signal Transduction; signal-transducer and activator of transcription; src Homology Domains; STAT1 Transcription Factor; STAT3 Transcription Factor; Tetracycline - pharmacology; Trans-Activators - metabolism; Transfection; localization; regulatable expression system; SHP-1; Protein tyrosine phosphatase; epidermal growth factor receptor; signal-transducer and activator of transcription
• ISSN: 0171-9335; 1618-1298
• DOI: 10.1078/S0171-9335(04)70029-1

The SH2 domain protein-tyrosine phosphatase SHP-1 has been shown earlier to bind to the epidermal growth factor receptor and to have the capacity for receptor dephosphorylation. New bi- and tricistronic expression vectors (pNRTIS-21 and pNRTIS-33, respectively) based on the tetracycline system were constructed and employed to generate stable cell lines with inducible expression of SHP-1. Inducible overexpression of SHP-1 in A431 cells led to attenuation of epidermal growth factor (EGF) receptor autophosphorylation and of EGF-induced DNA binding of ‘signal transducers and activators of transcription’ (STAT) 1 and 3. SHP-1 was localized in the cytoplasm with an enrichment in the perinuclear compartment. Association of SHP-1 with perinuclear structures may form the basis for a partial cofractionation with nuclei observed in different types of transfected cells and also with endogenous SHP-1 in U-937 cells. Treatment of SHP-1-overexpressing A431 cells or of HaCaT human keratinocytes expressing SHP-1 endogenously with the Ca
2+-ionophore A23187 resulted in partial nuclear accumulation of SHP-1. Thus, SHP-1 may interact with substrates or regulatory proteins in perinuclear or nuclear structures.