SHP-1 regulation of p62(DOK) tyrosine phosphorylation in macrophages

Karen L. Berg, Katherine A. Siminovitch, E. Richard Stanley

Research output: Contribution to journalArticle

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Abstract

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 SHIP-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.

Original languageEnglish (US)
Pages (from-to)35855-35865
Number of pages11
JournalJournal of Biological Chemistry
Volume274
Issue number50
DOIs
StatePublished - Dec 10 1999

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Phosphorylation
Macrophages
Macrophage Colony-Stimulating Factor
Tyrosine
Bone
Bone Marrow
Intercellular Signaling Peptides and Proteins
Association reactions
Cell signaling
HEK293 Cells
Phosphoproteins
Cell death
Cell Communication
Conformations
Catalytic Domain
Proteins
Cell Death
Modulation
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

SHP-1 regulation of p62(DOK) tyrosine phosphorylation in macrophages. / Berg, Karen L.; Siminovitch, Katherine A.; Stanley, E. Richard.

In: Journal of Biological Chemistry, Vol. 274, No. 50, 10.12.1999, p. 35855-35865.

Research output: Contribution to journalArticle

Berg, Karen L. ; Siminovitch, Katherine A. ; Stanley, E. Richard. / SHP-1 regulation of p62(DOK) tyrosine phosphorylation in macrophages. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 50. pp. 35855-35865.
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