Abstract
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 Ca2+-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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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3T3 Cells
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Animals
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Calcimycin / pharmacology
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Cell Line
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Cell Nucleus / metabolism*
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DNA, Complementary / metabolism
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DNA-Binding Proteins / metabolism*
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Enzyme Activation
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Epidermal Growth Factor / metabolism*
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ErbB Receptors / metabolism
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Humans
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Immunohistochemistry
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Intracellular Signaling Peptides and Proteins
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Ionophores / pharmacology
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Keratinocytes / metabolism
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Mice
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Protein Structure, Tertiary
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Protein Synthesis Inhibitors / pharmacology
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Protein Tyrosine Phosphatase, Non-Receptor Type 11
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Protein Tyrosine Phosphatase, Non-Receptor Type 6
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Protein Tyrosine Phosphatases / metabolism*
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SH2 Domain-Containing Protein Tyrosine Phosphatases
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STAT1 Transcription Factor
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STAT3 Transcription Factor
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Signal Transduction
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Tetracycline / pharmacology
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Trans-Activators / metabolism*
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Transfection
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src Homology Domains
Substances
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DNA, Complementary
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DNA-Binding Proteins
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Intracellular Signaling Peptides and Proteins
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Ionophores
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Protein Synthesis Inhibitors
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STAT1 Transcription Factor
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STAT1 protein, human
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STAT3 Transcription Factor
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STAT3 protein, human
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Stat1 protein, mouse
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Stat3 protein, mouse
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Trans-Activators
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Calcimycin
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Epidermal Growth Factor
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ErbB Receptors
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PTPN11 protein, human
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PTPN6 protein, human
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Protein Tyrosine Phosphatase, Non-Receptor Type 11
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Protein Tyrosine Phosphatase, Non-Receptor Type 6
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Protein Tyrosine Phosphatases
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Ptpn11 protein, mouse
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Ptpn6 protein, mouse
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SH2 Domain-Containing Protein Tyrosine Phosphatases
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Tetracycline