Characterization of a novel tyrosine phosphorylated 100-kDa protein that binds to SHP-2 and phosphatidylinositol 3'-kinase in myeloid cells.
Fms is a tyrosine kinase-containing receptor for macrophage colony-stimulating factor (M-CSF) that regulates survival, growth, and differentiation of cells along the monocyte/macrophage lineage. M-CSF stimulation of murine myeloid FDC-P1 cells expressing Fms resulted in the tyrosine phosphorylation of a number of signal transduction proteins, including an unidentified 100-kDa protein. This 100-kDa protein associated with the tyrosine phosphatase SHP-2 but not with the related phosphatase SHP-1. The kinetics of tyrosine phosphorylation of p100 and SHP-2 suggest that p100 may be a direct substrate of SHP-2. p100 bound directly to the SH2 domains of both SHP-2 and the p85 subunit of phosphatidylinositol 3'-kinase. The 100-kDa protein did not appear to bind directly to Fms, Ship, Cbl, Shc, or Grb2, although all of these proteins were coimmunoprecipitated with p85 after M-CSF stimulation. Association of p100 with SHP-2 and p85 did not require the major autophosphorylation sites on Fms nor binding of p85 to Fms. A tyrosine phosphorylated protein of 100 kDa also coprecipitated with SHP-2 from several other myeloid cell lines after M-CSF stimulation but was not seen in immunoprecipitates from Rat2 fibroblasts expressing Fms. Stimulation of FDC-P1 cells with additional cytokines also resulted in coprecipitation of a 100-kDa protein with SHP-2. p100 may therefore be a common component of the signaling pathways of cytokine receptors in myeloid cells.
Carlberg
K
K
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington, 98109-1024, USA. kcarlber@fhcrc.org
Rohrschneider
L R
LR
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D006224:Cricetinae; D005347:Fibroblasts; D051380:GRB2 Adaptor Protein; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins; D016173:Macrophage Colony-Stimulating Factor / Q000378:metabolism*; D051379:Mice; D008958:Models, Molecular; D008970:Molecular Weight; D050722:Oncogene Protein v-cbl; D019869:Phosphatidylinositol 3-Kinases; D000072183:Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D011485:Protein Binding; D021382:Protein Sorting Signals / Q000378:metabolism; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D051381:Rats; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000378:metabolism; D015682:Retroviridae Proteins, Oncogenic / Q000378:metabolism; D054593:SH2 Domain-Containing Protein Tyrosine Phosphatases; D055765:Shc Signaling Adaptor Proteins; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D014443:Tyrosine / Q000378:metabolism; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D051380:GRB2 Adaptor Protein; C495949:Grb2 protein, mouse; C495950:Grb2 protein, rat; D047908:Intracellular Signaling Peptides and Proteins; D050722:Oncogene Protein v-cbl; D021382:Protein Sorting Signals; D011506:Proteins; D015682:Retroviridae Proteins, Oncogenic; D055765:Shc Signaling Adaptor Proteins; C528034:Shc1 protein, mouse; C528035:Shc1 protein, rat; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D014443:Tyrosine; D016173:Macrophage Colony-Stimulating Factor; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D016186:Receptor, Macrophage Colony-Stimulating Factor; D010744:Phosphoric Monoester Hydrolases; C516724:PTPN11 protein, human; C509249:PTPN6 protein, human; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases; C516725:Ptpn11 protein, mouse; C516727:Ptpn11 protein, rat; C509248:Ptpn6 protein, mouse; C509250:Ptpn6 protein, rat; D054593:SH2 Domain-Containing Protein Tyrosine Phosphatases; C098706:INPPL1 protein, human; D000072183:Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases