Perinuclear Localization of the Protein-Tyrosine Phosphatase SHP-1 and Inhibition of Epidermal Growth Factor-Stimulated STAT1/3 Activation in A431 Cells

Eur J Cell Biol. 2000 Apr;79(4):261-71. doi: 10.1078/S0171-9335(04)70029-1.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Animals
  • Calcimycin / pharmacology
  • Cell Line
  • Cell Nucleus / metabolism*
  • DNA, Complementary / metabolism
  • DNA-Binding Proteins / metabolism*
  • Enzyme Activation
  • Epidermal Growth Factor / metabolism*
  • ErbB Receptors / metabolism
  • Humans
  • Immunohistochemistry
  • Intracellular Signaling Peptides and Proteins
  • Ionophores / pharmacology
  • Keratinocytes / metabolism
  • Mice
  • Protein Structure, Tertiary
  • Protein Synthesis Inhibitors / pharmacology
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6
  • Protein Tyrosine Phosphatases / metabolism*
  • SH2 Domain-Containing Protein Tyrosine Phosphatases
  • STAT1 Transcription Factor
  • STAT3 Transcription Factor
  • Signal Transduction
  • Tetracycline / pharmacology
  • Trans-Activators / metabolism*
  • Transfection
  • src Homology Domains

Substances

  • DNA, Complementary
  • DNA-Binding Proteins
  • Intracellular Signaling Peptides and Proteins
  • Ionophores
  • Protein Synthesis Inhibitors
  • STAT1 Transcription Factor
  • STAT1 protein, human
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • Stat1 protein, mouse
  • Stat3 protein, mouse
  • Trans-Activators
  • Calcimycin
  • Epidermal Growth Factor
  • ErbB Receptors
  • PTPN11 protein, human
  • PTPN6 protein, human
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6
  • Protein Tyrosine Phosphatases
  • Ptpn11 protein, mouse
  • Ptpn6 protein, mouse
  • SH2 Domain-Containing Protein Tyrosine Phosphatases
  • Tetracycline