• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcinvestThe Journal of Clinical InvestigationCurrent IssueArchiveSubscriptionAbout the Journal
J Clin Invest. Nov 15, 1998; 102(10): 1850–1859.
PMCID: PMC509135

Estrogen blocks M-CSF gene expression and osteoclast formation by regulating phosphorylation of Egr-1 and its interaction with Sp-1.

Abstract

Central to the pathogenesis of osteoporosis is the ability of estrogen deficiency to increase osteoclast formation by enhancing stromal cell production of the osteoclastogenic cytokine macrophage colony-stimulating factor (M-CSF). We report that stromal cells from ovariectomized mice exhibit increased casein kinase II-dependent phosphorylation of the nuclear protein Egr-1. Phosphorylated Egr-1 binds less avidly to the transcriptional activator Sp-1 and the resulting higher levels of free Sp-1 stimulate transactivation of the M-CSF gene. Estrogen replacement fails to block M-CSF mRNA expression and osteoclast formation in ovariectomized mice lacking Egr-1, confirming the critical role played by this transcription factor in mediating the antiosteoclastogenic effects of estrogen. Thus, by downregulating formation of a novel Egr-1/Sp-1 complex in stromal cells, estrogen deficiency results in enhanced levels of free Sp-1 and increased M-CSF gene expression and osteoclast formation.

Full Text

The Full Text of this article is available as a PDF (568K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Manolagas SC, Jilka RL. Bone marrow, cytokines, and bone remodeling. Emerging insights into the pathophysiology of osteoporosis. N Engl J Med. 1995 Feb 2;332(5):305–311. [PubMed]
  • Roodman GD. Advances in bone biology: the osteoclast. Endocr Rev. 1996 Aug;17(4):308–332. [PubMed]
  • Tanaka S, Takahashi N, Udagawa N, Tamura T, Akatsu T, Stanley ER, Kurokawa T, Suda T. Macrophage colony-stimulating factor is indispensable for both proliferation and differentiation of osteoclast progenitors. J Clin Invest. 1993 Jan;91(1):257–263. [PMC free article] [PubMed]
  • Yoshida H, Hayashi S, Kunisada T, Ogawa M, Nishikawa S, Okamura H, Sudo T, Shultz LD, 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. [PubMed]
  • Sarma U, Flanagan AM. Macrophage colony-stimulating factor induces substantial osteoclast generation and bone resorption in human bone marrow cultures. Blood. 1996 Oct 1;88(7):2531–2540. [PubMed]
  • Sherman ML, Weber BL, Datta R, Kufe DW. Transcriptional and posttranscriptional regulation of macrophage-specific colony stimulating factor gene expression by tumor necrosis factor. Involvement of arachidonic acid metabolites. J Clin Invest. 1990 Feb;85(2):442–447. [PMC free article] [PubMed]
  • Falkenburg JH, Harrington MA, de Paus RA, Walsh WK, Daub R, Landegent JE, Broxmeyer HE. Differential transcriptional and posttranscriptional regulation of gene expression of the colony-stimulating factors by interleukin-1 and fetal bovine serum in murine fibroblasts. Blood. 1991 Aug 1;78(3):658–665. [PubMed]
  • Pfeilschifter J, Chenu C, Bird A, Mundy GR, Roodman GD. 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. [PubMed]
  • Kimble RB, Srivastava S, Ross FP, Matayoshi A, Pacifici R. Estrogen deficiency increases the ability of stromal cells to support murine osteoclastogenesis via an interleukin-1and tumor necrosis factor-mediated stimulation of macrophage colony-stimulating factor production. J Biol Chem. 1996 Nov 15;271(46):28890–28897. [PubMed]
  • Harrington MA, Edenberg HJ, Saxman S, Pedigo LM, Daub R, Broxmeyer HE. Cloning and characterization of the murine promoter for the colony-stimulating factor-1-encoding gene. Gene. 1991 Jun 30;102(2):165–170. [PubMed]
  • Cao XM, Koski RA, Gashler A, McKiernan M, Morris CF, Gaffney R, Hay RV, Sukhatme VP. Identification and characterization of the Egr-1 gene product, a DNA-binding zinc finger protein induced by differentiation and growth signals. Mol Cell Biol. 1990 May;10(5):1931–1939. [PMC free article] [PubMed]
  • Milbrandt J. A nerve growth factor-induced gene encodes a possible transcriptional regulatory factor. Science. 1987 Nov 6;238(4828):797–799. [PubMed]
  • Kadonaga JT, Carner KR, Masiarz FR, Tjian R. Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain. Cell. 1987 Dec 24;51(6):1079–1090. [PubMed]
  • Hagen G, Müller S, Beato M, Suske G. Sp1-mediated transcriptional activation is repressed by Sp3. EMBO J. 1994 Aug 15;13(16):3843–3851. [PMC free article] [PubMed]
  • Jain N, Mahendran R, Philp R, Guy GR, Tan YH, Cao X. Casein kinase II associates with Egr-1 and acts as a negative modulator of its DNA binding and transcription activities in NIH 3T3 cells. J Biol Chem. 1996 Jun 7;271(23):13530–13536. [PubMed]
  • Allende JE, Allende CC. Protein kinases. 4. Protein kinase CK2: an enzyme with multiple substrates and a puzzling regulation. FASEB J. 1995 Mar;9(5):313–323. [PubMed]
  • Kitazawa R, Kimble RB, Vannice JL, Kung VT, Pacifici R. Interleukin-1 receptor antagonist and tumor necrosis factor binding protein decrease osteoclast formation and bone resorption in ovariectomized mice. J Clin Invest. 1994 Dec;94(6):2397–2406. [PMC free article] [PubMed]
  • Lee SL, Tourtellotte LC, Wesselschmidt RL, Milbrandt J. Growth and differentiation proceeds normally in cells deficient in the immediate early gene NGFI-A. J Biol Chem. 1995 Apr 28;270(17):9971–9977. [PubMed]
  • Lee SL, Sadovsky Y, Swirnoff AH, Polish JA, Goda P, Gavrilina G, Milbrandt J. Luteinizing hormone deficiency and female infertility in mice lacking the transcription factor NGFI-A (Egr-1). Science. 1996 Aug 30;273(5279):1219–1221. [PubMed]
  • Godambe SA, Chaplin DD, Takova T, Read LM, Bellone CJ. A novel cis-acting element required for lipopolysaccharide-induced transcription of the murine interleukin-1 beta gene. Mol Cell Biol. 1995 Jan;15(1):112–119. [PMC free article] [PubMed]
  • Hall CV, Jacob PE, Ringold GM, Lee F. Expression and regulation of Escherichia coli lacZ gene fusions in mammalian cells. J Mol Appl Genet. 1983;2(1):101–109. [PubMed]
  • Schneider I. Cell lines derived from late embryonic stages of Drosophila melanogaster. J Embryol Exp Morphol. 1972 Apr;27(2):353–365. [PubMed]
  • Maniatis T, Goodbourn S, Fischer JA. Regulation of inducible and tissue-specific gene expression. Science. 1987 Jun 5;236(4806):1237–1245. [PubMed]
  • Andrews NC, Faller DV. A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells. Nucleic Acids Res. 1991 May 11;19(9):2499–2499. [PMC free article] [PubMed]
  • Tetsuka T, Srivastava SK, Morrison AR. Tyrosine kinase inhibitors, genistein and herbimycin A, do not block interleukin-1 beta-induced activation of NF-kappa B in rat mesangial cells. Biochem Biophys Res Commun. 1996 Jan 26;218(3):808–812. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PMC free article] [PubMed]
  • Walton GM, Bertics PJ, Hudson LG, Vedvick TS, Gill GN. A three-step purification procedure for protein kinase C: characterization of the purified enzyme. Anal Biochem. 1987 Mar;161(2):425–437. [PubMed]
  • Heller-Harrison RA, Czech MP. Enhanced casein kinase II activity in COS-1 cells upon overexpression of either its catalytic or noncatalytic subunit. J Biol Chem. 1991 Aug 5;266(22):14435–14439. [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • Courey AJ, Tjian R. Analysis of Sp1 in vivo reveals multiple transcriptional domains, including a novel glutamine-rich activation motif. Cell. 1988 Dec 2;55(5):887–898. [PubMed]
  • McNabb DS, Guarente L. Genetic and biochemical probes for protein-protein interactions. Curr Opin Biotechnol. 1996 Oct;7(5):554–559. [PubMed]
  • Bai C, Elledge SJ. Gene identification using the yeast two-hybrid system. Methods Enzymol. 1996;273:331–347. [PubMed]
  • Luban J, Goff SP. The yeast two-hybrid system for studying protein-protein interactions. Curr Opin Biotechnol. 1995 Feb;6(1):59–64. [PubMed]
  • Day ML, Fahrner TJ, Aykent S, Milbrandt J. The zinc finger protein NGFI-A exists in both nuclear and cytoplasmic forms in nerve growth factor-stimulated PC12 cells. J Biol Chem. 1990 Sep 5;265(25):15253–15260. [PubMed]
  • Kuenzel EA, Krebs EG. A synthetic peptide substrate specific for casein kinase II. Proc Natl Acad Sci U S A. 1985 Feb;82(3):737–741. [PMC free article] [PubMed]
  • Jilka RL, Hangoc G, Girasole G, Passeri G, Williams DC, Abrams JS, Boyce B, Broxmeyer H, Manolagas SC. Increased osteoclast development after estrogen loss: mediation by interleukin-6. Science. 1992 Jul 3;257(5066):88–91. [PubMed]
  • Skerka C, Decker EL, Zipfel PF. A regulatory element in the human interleukin 2 gene promoter is a binding site for the zinc finger proteins Sp1 and EGR-1. J Biol Chem. 1995 Sep 22;270(38):22500–22506. [PubMed]
  • Krämer B, Meichle A, Hensel G, Charnay P, Krönke M. Characterization of an Krox-24/Egr-1-responsive element in the human tumor necrosis factor promoter. Biochim Biophys Acta. 1994 Oct 18;1219(2):413–421. [PubMed]
  • Dennig J, Hagen G, Beato M, Suske G. Members of the Sp transcription factor family control transcription from the uteroglobin promoter. J Biol Chem. 1995 May 26;270(21):12737–12744. [PubMed]
  • Ackerman SL, Minden AG, Williams GT, Bobonis C, Yeung CY. Functional significance of an overlapping consensus binding motif for Sp1 and Zif268 in the murine adenosine deaminase gene promoter. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7523–7527. [PMC free article] [PubMed]
  • Harrington MA, Konicek B, Song A, Xia XL, Fredericks WJ, Rauscher FJ., 3rd Inhibition of colony-stimulating factor-1 promoter activity by the product of the Wilms' tumor locus. J Biol Chem. 1993 Oct 5;268(28):21271–21275. [PubMed]
  • Ebert SN, Wong DL. Differential activation of the rat phenylethanolamine N-methyltransferase gene by Sp1 and Egr-1. J Biol Chem. 1995 Jul 21;270(29):17299–17305. [PubMed]
  • Gruss HJ, Duyster J, Herrmann F. Structural and biological features of the TNF receptor and TNF ligand superfamilies: interactive signals in the pathobiology of Hodgkin's disease. Ann Oncol. 1996;7 (Suppl 4):19–26. [PubMed]
  • Martin MU, Falk W. The interleukin-1 receptor complex and interleukin-1 signal transduction. Eur Cytokine Netw. 1997 Mar;8(1):5–17. [PubMed]
  • Kimble RB, Vannice JL, Bloedow DC, Thompson RC, Hopfer W, Kung VT, Brownfield C, Pacifici R. Interleukin-1 receptor antagonist decreases bone loss and bone resorption in ovariectomized rats. J Clin Invest. 1994 May;93(5):1959–1967. [PMC free article] [PubMed]
  • Kimble RB, Matayoshi AB, Vannice JL, Kung VT, Williams C, Pacifici R. Simultaneous block of interleukin-1 and tumor necrosis factor is required to completely prevent bone loss in the early postovariectomy period. Endocrinology. 1995 Jul;136(7):3054–3061. [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...