Abstract
Colony-stimulating factor-1 (CSF-1) stimulates motility and cytoplasmic spreading in mature osteoclasts. Therefore, we examined the cellular events and intracellular signaling pathways that accompany CSF-1-induced spreading in normal osteoclasts. To explore the role c-src plays in these processes, we also studied osteoclasts prepared from animals with targeted disruption of the src gene. In normal osteoclasts, CSF-1 treatment induces rapid cytoplasmic spreading, with redistribution of F-actin from a well-delineated central attachment ring to the periphery of the cell. CSF-1 increases membrane phosphotyrosine staining in osteoclasts and induces the phosphorylation of several cellular proteins in cultured, osteoclast-like cells, including c-fms, c-src, and an 85-kD Grb2-binding protein. Src kinase activity is increased threefold after CSF-1 treatment. In src- cells, no attachment ring is present, and CSF-1 fails to induce spreading or a change in the pattern of F-actin distribution. Although c-fms becomes phosphorylated after CSF-1 treatment, the 85-kD protein is significantly less phosphorylated in src- osteoclast-like cells. These results indicate that c-src is critical for the normal cytoskeletal architecture of the osteoclast, and, in its absence, the spreading response induced by CSF-1 is abrogated, and downstream signaling from c-fms is altered.
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- Akatsu T., Tamura T., Takahashi N., Udagawa N., Tanaka S., Sasaki T., Yamaguchi A., Nagata N., Suda T. Preparation and characterization of a mouse osteoclast-like multinucleated cell population. J Bone Miner Res. 1992 Nov;7(11):1297–1306. doi: 10.1002/jbmr.5650071109. [DOI] [PubMed] [Google Scholar]
- Alonso G., Koegl M., Mazurenko N., Courtneidge S. A. Sequence requirements for binding of Src family tyrosine kinases to activated growth factor receptors. J Biol Chem. 1995 Apr 28;270(17):9840–9848. doi: 10.1074/jbc.270.17.9840. [DOI] [PubMed] [Google Scholar]
- Ambros V. R., Chen L. B., Buchanan J. M. Surface ruffles as markers for studies of cell transformation by Rous sarcoma virus. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3144–3148. doi: 10.1073/pnas.72.8.3144. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boocock C. A., Jones G. E., Stanley E. R., Pollard J. W. Colony-stimulating factor-1 induces rapid behavioural responses in the mouse macrophage cell line, BAC1.2F5. J Cell Sci. 1989 Jul;93(Pt 3):447–456. doi: 10.1242/jcs.93.3.447. [DOI] [PubMed] [Google Scholar]
- Boyce B. F., Yoneda T., Lowe C., Soriano P., Mundy G. R. Requirement of pp60c-src expression for osteoclasts to form ruffled borders and resorb bone in mice. J Clin Invest. 1992 Oct;90(4):1622–1627. doi: 10.1172/JCI116032. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Courtneidge S. A., Dhand R., Pilat D., Twamley G. M., Waterfield M. D., Roussel M. F. Activation of Src family kinases by colony stimulating factor-1, and their association with its receptor. EMBO J. 1993 Mar;12(3):943–950. doi: 10.1002/j.1460-2075.1993.tb05735.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- David-Pfeuty T., Singer S. J. Altered distributions of the cytoskeletal proteins vinculin and alpha-actinin in cultured fibroblasts transformed by Rous sarcoma virus. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6687–6691. doi: 10.1073/pnas.77.11.6687. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elford P. R., Felix R., Cecchini M., Trechsel U., Fleisch H. Murine osteoblastlike cells and the osteogenic cell MC3T3-E1 release a macrophage colony-stimulating activity in culture. Calcif Tissue Int. 1987 Sep;41(3):151–156. doi: 10.1007/BF02563795. [DOI] [PubMed] [Google Scholar]
- Elias J. A., Tang W., Horowitz M. C. Cytokine and hormonal stimulation of human osteosarcoma interleukin-11 production. Endocrinology. 1995 Feb;136(2):489–498. doi: 10.1210/endo.136.2.7835281. [DOI] [PubMed] [Google Scholar]
- Felice G. R., Eason P., Nermut M. V., Kellie S. pp60v-src association with the cytoskeleton induces actin reorganization without affecting polymerization status. Eur J Cell Biol. 1990 Jun;52(1):47–59. [PubMed] [Google Scholar]
- Felix R., Cecchini M. G., Fleisch H. Macrophage colony stimulating factor restores in vivo bone resorption in the op/op osteopetrotic mouse. Endocrinology. 1990 Nov;127(5):2592–2594. doi: 10.1210/endo-127-5-2592. [DOI] [PubMed] [Google Scholar]
- Felix R., Elford P. R., Stoercklé C., Cecchini M., Wetterwald A., Trechsel U., Fleisch H., Stadler B. M. Production of hemopoietic growth factors by bone tissue and bone cells in culture. J Bone Miner Res. 1988 Feb;3(1):27–36. doi: 10.1002/jbmr.5650030106. [DOI] [PubMed] [Google Scholar]
- Felix R., Fleisch H., Elford P. R. Bone-resorbing cytokines enhance release of macrophage colony-stimulating activity by the osteoblastic cell MC3T3-E1. Calcif Tissue Int. 1989 May;44(5):356–360. doi: 10.1007/BF02556317. [DOI] [PubMed] [Google Scholar]
- Fuller K., Owens J. M., Jagger C. J., Wilson A., Moss R., Chambers T. J. Macrophage colony-stimulating factor stimulates survival and chemotactic behavior in isolated osteoclasts. J Exp Med. 1993 Nov 1;178(5):1733–1744. doi: 10.1084/jem.178.5.1733. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Girasole G., Passeri G., Jilka R. L., Manolagas S. C. Interleukin-11: a new cytokine critical for osteoclast development. J Clin Invest. 1994 Apr;93(4):1516–1524. doi: 10.1172/JCI117130. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hofstetter W., Wetterwald A., Cecchini M. C., Felix R., Fleisch H., Mueller C. Detection of transcripts for the receptor for macrophage colony-stimulating factor, c-fms, in murine osteoclasts. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9637–9641. doi: 10.1073/pnas.89.20.9637. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horne W. C., Neff L., Chatterjee D., Lomri A., Levy J. B., Baron R. Osteoclasts express high levels of pp60c-src in association with intracellular membranes. J Cell Biol. 1992 Nov;119(4):1003–1013. doi: 10.1083/jcb.119.4.1003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kanehisa J., Heersche J. N. Osteoclastic bone resorption: in vitro analysis of the rate of resorption and migration of individual osteoclasts. Bone. 1988;9(2):73–79. doi: 10.1016/8756-3282(88)90106-8. [DOI] [PubMed] [Google Scholar]
- Kaplan K. B., Swedlow J. R., Morgan D. O., Varmus H. E. c-Src enhances the spreading of src-/- fibroblasts on fibronectin by a kinase-independent mechanism. Genes Dev. 1995 Jun 15;9(12):1505–1517. doi: 10.1101/gad.9.12.1505. [DOI] [PubMed] [Google Scholar]
- Keeting P. E., Rifas L., Harris S. A., Colvard D. S., Spelsberg T. C., Peck W. A., Riggs B. L. Evidence for interleukin-1 beta production by cultured normal human osteoblast-like cells. J Bone Miner Res. 1991 Aug;6(8):827–833. doi: 10.1002/jbmr.5650060807. [DOI] [PubMed] [Google Scholar]
- Klint P., Kanda S., Claesson-Welsh L. Shc and a novel 89-kDa component couple to the Grb2-Sos complex in fibroblast growth factor-2-stimulated cells. J Biol Chem. 1995 Oct 6;270(40):23337–23344. doi: 10.1074/jbc.270.40.23337. [DOI] [PubMed] [Google Scholar]
- Kogan S. C., Doherty M., Gitschier J. An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences. Application to hemophilia A. N Engl J Med. 1987 Oct 15;317(16):985–990. doi: 10.1056/NEJM198710153171603. [DOI] [PubMed] [Google Scholar]
- Kouhara H., Hadari Y. R., Spivak-Kroizman T., Schilling J., Bar-Sagi D., Lax I., Schlessinger J. A lipid-anchored Grb2-binding protein that links FGF-receptor activation to the Ras/MAPK signaling pathway. Cell. 1997 May 30;89(5):693–702. doi: 10.1016/s0092-8674(00)80252-4. [DOI] [PubMed] [Google Scholar]
- Kurihara N., Bertolini D., Suda T., Akiyama Y., Roodman G. D. IL-6 stimulates osteoclast-like multinucleated cell formation in long term human marrow cultures by inducing IL-1 release. J Immunol. 1990 Jun 1;144(11):4226–4230. [PubMed] [Google Scholar]
- Lassar A. B., Buskin J. N., Lockshon D., Davis R. L., Apone S., Hauschka S. D., Weintraub H. MyoD is a sequence-specific DNA binding protein requiring a region of myc homology to bind to the muscle creatine kinase enhancer. Cell. 1989 Sep 8;58(5):823–831. doi: 10.1016/0092-8674(89)90935-5. [DOI] [PubMed] [Google Scholar]
- Lee A. W., Nienhuis A. W. Mechanism of kinase activation in the receptor for colony-stimulating factor 1. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7270–7274. doi: 10.1073/pnas.87.18.7270. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levy J. B., Brugge J. S. Biological and biochemical properties of the c-src+ gene product overexpressed in chicken embryo fibroblasts. Mol Cell Biol. 1989 Aug;9(8):3332–3341. doi: 10.1128/mcb.9.8.3332. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lioubin M. N., Myles G. M., Carlberg K., Bowtell D., Rohrschneider L. R. Shc, Grb2, Sos1, and a 150-kilodalton tyrosine-phosphorylated protein form complexes with Fms in hematopoietic cells. Mol Cell Biol. 1994 Sep;14(9):5682–5691. doi: 10.1128/mcb.14.9.5682. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lowe C., Yoneda T., Boyce B. F., Chen H., Mundy G. R., Soriano P. Osteopetrosis in Src-deficient mice is due to an autonomous defect of osteoclasts. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4485–4489. doi: 10.1073/pnas.90.10.4485. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lowenstein E. J., Daly R. J., Batzer A. G., Li W., Margolis B., Lammers R., Ullrich A., Skolnik E. Y., Bar-Sagi D., Schlessinger J. The SH2 and SH3 domain-containing protein GRB2 links receptor tyrosine kinases to ras signaling. Cell. 1992 Aug 7;70(3):431–442. doi: 10.1016/0092-8674(92)90167-b. [DOI] [PubMed] [Google Scholar]
- Meenakshi T., Ross F. P., Martin J., Teitelbaum S. L. 1,25-Dihydroxyvitamin D3 and macrophage colony-stimulating factor-1 synergistically phosphorylate talin. J Cell Biochem. 1993 Oct;53(2):145–155. doi: 10.1002/jcb.240530207. [DOI] [PubMed] [Google Scholar]
- Mohammadi M., Dikic I., Sorokin A., Burgess W. H., Jaye M., Schlessinger J. Identification of six novel autophosphorylation sites on fibroblast growth factor receptor 1 and elucidation of their importance in receptor activation and signal transduction. Mol Cell Biol. 1996 Mar;16(3):977–989. doi: 10.1128/mcb.16.3.977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Owens J., Chambers T. J. Macrophage colony-stimulating factor (M-CSF) induces migration in osteoclasts in vitro. Biochem Biophys Res Commun. 1993 Sep 30;195(3):1401–1407. doi: 10.1006/bbrc.1993.2199. [DOI] [PubMed] [Google Scholar]
- Parsons J. T., Weber M. J. Genetics of src: structure and functional organization of a protein tyrosine kinase. Curr Top Microbiol Immunol. 1989;147:79–127. doi: 10.1007/978-3-642-74697-0_3. [DOI] [PubMed] [Google Scholar]
- Pfeilschifter J., Chenu C., Bird A., Mundy G. R., Roodman G. D. Interleukin-1 and tumor necrosis factor stimulate the formation of human osteoclastlike cells in vitro. J Bone Miner Res. 1989 Feb;4(1):113–118. doi: 10.1002/jbmr.5650040116. [DOI] [PubMed] [Google Scholar]
- Roodman G. D. Advances in bone biology: the osteoclast. Endocr Rev. 1996 Aug;17(4):308–332. doi: 10.1210/edrv-17-4-308. [DOI] [PubMed] [Google Scholar]
- Sengupta A., Liu W. K., Yeung Y. G., Yeung D. C., Frackelton A. R., Jr, Stanley E. R. Identification and subcellular localization of proteins that are rapidly phosphorylated in tyrosine in response to colony-stimulating factor 1. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8062–8066. doi: 10.1073/pnas.85.21.8062. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Soriano P., Montgomery C., Geske R., Bradley A. Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice. Cell. 1991 Feb 22;64(4):693–702. doi: 10.1016/0092-8674(91)90499-o. [DOI] [PubMed] [Google Scholar]
- Su Y., Chakraborty M., Nathanson M. H., Baron R. Differential effects of the 3',5'-cyclic adenosine monophosphate and protein kinase C pathways on the response of isolated rat osteoclasts to calcitonin. Endocrinology. 1992 Sep;131(3):1497–1502. doi: 10.1210/endo.131.3.1324163. [DOI] [PubMed] [Google Scholar]
- Tanaka S., Amling M., Neff L., Peyman A., Uhlmann E., Levy J. B., Baron R. c-Cbl is downstream of c-Src in a signalling pathway necessary for bone resorption. Nature. 1996 Oct 10;383(6600):528–531. doi: 10.1038/383528a0. [DOI] [PubMed] [Google Scholar]
- Thomas S. M., Soriano P., Imamoto A. Specific and redundant roles of Src and Fyn in organizing the cytoskeleton. Nature. 1995 Jul 20;376(6537):267–271. doi: 10.1038/376267a0. [DOI] [PubMed] [Google Scholar]
- Wang J. M., Griffin J. D., Rambaldi A., Chen Z. G., Mantovani A. Induction of monocyte migration by recombinant macrophage colony-stimulating factor. J Immunol. 1988 Jul 15;141(2):575–579. [PubMed] [Google Scholar]
- Wang Y., Yeung Y. G., Langdon W. Y., Stanley E. R. c-Cbl is transiently tyrosine-phosphorylated, ubiquitinated, and membrane-targeted following CSF-1 stimulation of macrophages. J Biol Chem. 1996 Jan 5;271(1):17–20. doi: 10.1074/jbc.271.1.17. [DOI] [PubMed] [Google Scholar]
- Weir E. C., Horowitz M. C., Baron R., Centrella M., Kacinski B. M., Insogna K. L. Macrophage colony-stimulating factor release and receptor expression in bone cells. J Bone Miner Res. 1993 Dec;8(12):1507–1518. doi: 10.1002/jbmr.5650081214. [DOI] [PubMed] [Google Scholar]
- Weir E. C., Insogna K. L., Horowitz M. C. Osteoblast-like cells secrete granulocyte-macrophage colony-stimulating factor in response to parathyroid hormone and lipopolysaccharide. Endocrinology. 1989 Feb;124(2):899–904. doi: 10.1210/endo-124-2-899. [DOI] [PubMed] [Google Scholar]
- Wiktor-Jedrzejczak W., Bartocci A., Ferrante A. W., Jr, Ahmed-Ansari A., Sell K. W., Pollard J. W., Stanley E. R. Total absence of colony-stimulating factor 1 in the macrophage-deficient osteopetrotic (op/op) mouse. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4828–4832. doi: 10.1073/pnas.87.12.4828. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yoshida H., Hayashi S., Kunisada T., Ogawa M., Nishikawa S., Okamura H., Sudo T., Shultz L. D., Nishikawa S. The murine mutation osteopetrosis is in the coding region of the macrophage colony stimulating factor gene. Nature. 1990 May 31;345(6274):442–444. doi: 10.1038/345442a0. [DOI] [PubMed] [Google Scholar]
- van der Geer P., Hunter T. Mutation of Tyr697, a GRB2-binding site, and Tyr721, a PI 3-kinase binding site, abrogates signal transduction by the murine CSF-1 receptor expressed in Rat-2 fibroblasts. EMBO J. 1993 Dec 15;12(13):5161–5172. doi: 10.1002/j.1460-2075.1993.tb06211.x. [DOI] [PMC free article] [PubMed] [Google Scholar]