Receptor activator of NF-kappa B ligand stimulates recruitment of SHP-1 to the complex containing TNFR-associated factor 6 that regulates osteoclastogenesis.
Receptor activator of NF-kappaB ligand (RANKL) is essential for differentiation and function of osteoclasts. The negative signaling pathways downstream of RANKL are not well characterized. By retroviral transduction of RAW264.7 cells with a dominant negative Src homology 2 domain-containing phosphatase-1 (SHP-1)(C453S), we studied the role of tyrosine phosphatase SHP-1 in ... RANKL-induced osteoclastogenesis. Over-expression of SHP-1(C453S) significantly enhanced the number of tartrate-resistant acid phosphatase-positive multinuclear osteoclast-like cells in response to RANKL in a dose-dependent manner. RANKL induced the recruitment of SHP-1 to a complex containing TNFR-associated factor (TRAF)6. GST pull down experiments indicated that the association of SHP-1 with TRAF6 is mediated by SHP-1 lacking the two Src homology 2 domains. RANKL-stimulated IkappaB-alpha phosphorylation, IkappaB-alpha degradation and DNA binding ability of NF-kappaB were increased after over-expression of SHP-1(C453S). However, RANKL-induced phosphorylation of mitogen-activated protein kinases, extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 mitogen-activated protein kinase, was unchanged. In addition, SHP-1 regulated RANKL-stimulated tyrosine phosphorylation of p85 subunit of phosphatidylinositol 3 kinase and the phosphorylation of Akt. Increased numbers of osteoclasts contribute to severe osteopenia in Me(v)/Me(v) mice due to mutation of SHP-1. Like RAW264.7 cells expressing SHP-1(C453S), the bone marrow macrophages of Me(v)/Me(v) mice generated much more osteoclast-like cells than that of littermate controls in response to RANKL. Furthermore compared with controls, RANKL induces enhanced association of TRAF6 and RANK in both RAW264.7 cells expressing SHP-1(C453S) and bone marrow macrophages from Me(v)/Me(v) mice. Therefore, SHP-1 plays a role in signals downstream of RANKL by recruitment to the complex containing TRAF6 and these observations may help to understand the mechanism of osteoporosis in Me(v)/Me(v) mice.
Mesh Terms:
Animals, Carrier Proteins, Cell Differentiation, Cell Line, Cysteine, Genetic Vectors, Glycoproteins, Intracellular Signaling Peptides and Proteins, Ligands, Macrophages, Membrane Glycoproteins, Mice, Mice, Inbred C57BL, Mice, Mutant Strains, NF-kappa B, Osteoclasts, Osteoprotegerin, Phosphatidylinositol 3-Kinases, Phosphorylation, Protein Phosphatase 1, Protein Subunits, Protein Transport, Protein Tyrosine Phosphatase, Non-Receptor Type 6, Protein Tyrosine Phosphatases, Protein-Serine-Threonine Kinases, Proteins, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-akt, RANK Ligand, Receptor Activator of Nuclear Factor-kappa B, Receptors, Cytoplasmic and Nuclear, Receptors, Tumor Necrosis Factor, Retroviridae, Serine, TNF Receptor-Associated Factor 6, Transduction, Genetic, Tyrosine, Up-Regulation, src Homology Domains
Animals, Carrier Proteins, Cell Differentiation, Cell Line, Cysteine, Genetic Vectors, Glycoproteins, Intracellular Signaling Peptides and Proteins, Ligands, Macrophages, Membrane Glycoproteins, Mice, Mice, Inbred C57BL, Mice, Mutant Strains, NF-kappa B, Osteoclasts, Osteoprotegerin, Phosphatidylinositol 3-Kinases, Phosphorylation, Protein Phosphatase 1, Protein Subunits, Protein Transport, Protein Tyrosine Phosphatase, Non-Receptor Type 6, Protein Tyrosine Phosphatases, Protein-Serine-Threonine Kinases, Proteins, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-akt, RANK Ligand, Receptor Activator of Nuclear Factor-kappa B, Receptors, Cytoplasmic and Nuclear, Receptors, Tumor Necrosis Factor, Retroviridae, Serine, TNF Receptor-Associated Factor 6, Transduction, Genetic, Tyrosine, Up-Regulation, src Homology Domains
J. Immunol.
Date: Oct. 01, 2003
PubMed ID: 14500659
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