SHP-1 regulation of p62(DOK) tyrosine phosphorylation in macrophages.
SHP-1 plays key roles in the modulation of hematopoietic cell signaling. To ascertain the impact of SHP-1 on colony-stimulating factor-1 (CSF-1)-mediated survival and proliferative signaling, we compared the CSF-1 responses of primary bone marrow macrophages (BMM) from wild-type and SHP-1-deficient motheaten (me/me) mice. CSF-1-induced protein tyrosine phosphorylation levels were similar in wild-type and me/me BMM, except for the constitutive hyperphosphorylation of a 62-kDa phosphoprotein (pp62) in me/me macrophages. pp62 was identified as the RASGAP-associated p62(DOK) and was shown to be the major CSF-1R-associated tyrosine-phosphorylated protein in CSF-1-treated BMM. p62(DOK) was found to be constitutively associated with SHP-1 in BMM and in 293T cells, co-expressing p62(dok) and either wild-type or catalytically inert SHP-1 (SHP-1 C453S). In both cell types, the interaction of SHP-1 with p62(DOK) occurred independently of p62(DOK) tyrosine phosphorylation, but only the tyrosine-phosphorylated p62(DOK) was bound by SHP-1 C453S in a far Western analysis. These findings suggest a constitutive association of SHP-1 and p62(DOK) that is either conformation-dependent or indirect as well as a direct, inducible association of the SHP-1 catalytic domain with tyrosine-phosphorylated p62(DOK). p62(DOK) hyperphosphorylation is not associated with altered CSF-1-induced RAS signaling or proliferation. However, whereas wild-type macrophages undergo cell death following CSF-1 removal, me/me macrophages exhibit prolonged survival in the absence of growth factor. Thus, p62(DOK) is a major SHP-1 substrate whose tyrosine phosphorylation correlates with growth factor-independent survival in macrophages.
Berg
K L
KL
Department of Developmental Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Siminovitch
K A
KA
Stanley
E R
ER
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002460:Cell Line; D002478:Cells, Cultured; D004268:DNA-Binding Proteins; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008817:Mice, Mutant Strains; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D054593:SH2 Domain-Containing Protein Tyrosine Phosphatases; D014162:Transfection; D018909:src Homology Domains
D004268:DNA-Binding Proteins; C489560:DOK1 protein, human; C489561:Dok1 protein, mouse; C074561:GAP-associated protein p62; D047908:Intracellular Signaling Peptides and Proteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D019000:Phosphotyrosine; D016173:Macrophage Colony-Stimulating Factor; D005982:Glutathione Transferase; 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; C509248:Ptpn6 protein, mouse; D054593:SH2 Domain-Containing Protein Tyrosine Phosphatases