title: M-CSF induces the stable interaction of cFms with alphaVbeta3 integrin in osteoclasts.
abstract: The macrophage colony stimulating factor receptor (cFms) and alpha(V)beta(3) integrin are both abundantly expressed and play critical roles in the differentiation, survival and migration of osteoclasts. We have previously demonstrated that cross-talk between cFms- and alpha(V)beta(3)-mediated signaling pathways regulated the cytoskeletal organization required for osteoclast migration. To investigate the nature of interaction between the two receptors, we sequentially used anion-exchange chromatography and immunoprecipitation to purify alpha(V)beta(3)-associated protein complexes. We have demonstrated that cFms stably associated with alpha(V)beta(3) in osteoclasts during adhesion, and that the association was induced by macrophage colony stimulating factor (M-CSF) stimulation. However, the kinetics of association of alpha(V)beta(3) and cFms did not correlate with the kinetics of tyrosine phosphorylation of cFms. Instead, maximally observed alpha(V)beta(3)/cFms association was after the peak of cFms tyrosine phosphorylation and correlated inversely with the total amount of cFms remaining. Furthermore, the complex containing cFms and alpha(V)beta(3) also contained a number of other signaling molecules including Pyk2, p130(Cas) and c-Cbl, known downstream regulators of the integrin-mediated signaling pathways in osteoclasts. In the presence of M-CSF, co-localization of alpha(V)beta(3) integrin and cFms was identified in the podosomal actin ring of the osteoclast during adhesion on glass. Interestingly, co-localization of both receptors was not found in the sealing zone, but in punctate structures associated with adhesion- or transcytosis-like structures in osteoclasts on bone. Taken together, we suggest that the association of alpha(V)beta(3) and cFms could be the result of signaling following tyrosine phosphorylation of cFms. The recruitment of cFms to alpha(V)beta(3) integrin may be an integral part of a larger signaling complex via which both of adhesion- and growth factor receptors coordinately regulate osteoclast adhesion, motility and membrane trafficking.
authors.affiliation: Department of Molecular Endocrinology & Bone Biology, Merck Research Laboratories, West Point, PA 19486, USA. c.elsegood@unimelb.edu.au
authors.forename: Caryn L
authors.initials: CL
authors.lastname: Elsegood
authors.forename: Ya
authors.initials: Y
authors.lastname: Zhuo
authors.forename: Gregg A
authors.initials: GA
authors.lastname: Wesolowski
authors.forename: John A
authors.initials: JA
authors.lastname: Hamilton
authors.forename: Gideon A
authors.initials: GA
authors.lastname: Rodan
authors.forename: Le T
authors.initials: LT
authors.lastname: Duong
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D050719:Crk-Associated Substrate Protein / Q000378:metabolism; D047468:Immunoprecipitation; D039302:Integrin alphaVbeta3 / Q000378:metabolism*; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology / Q000502:physiology*; D051379:Mice; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism*; D010766:Phosphorylation; D050721:Proto-Oncogene Proteins c-cbl / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology
chemical_list: D050719:Crk-Associated Substrate Protein; D039302:Integrin alphaVbeta3; D016173:Macrophage Colony-Stimulating Factor; D050721:Proto-Oncogene Proteins c-cbl; D020794:Receptor Protein-Tyrosine Kinases; D016186:Receptor, Macrophage Colony-Stimulating Factor