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Mol Med. 1996 November; 2(6): 766–773.
PMCID: PMC2230139

The beta c component of the granulocyte-macrophage colony-stimulating factor (GM-CSF)/interleukin 3 (IL-3)/IL-5 receptor interacts with a hybrid GM-CSF/erythropoietin receptor to influence proliferation and beta-globin mRNA expression.

Abstract

BACKGROUND: The interaction of different members of the hematopoietic growth factor receptor family may be relevant to the increased proliferation and the failure of differentiation that characterizes the myeloid leukemias. We recently demonstrated that a chimeric receptor (GMER) that is composed of the extracellular and transmembrane domains of the human granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor alpha-chain (GMR alpha) and the cytoplasmic domain of the murine erythropoietin receptor mEpoR binds hGM-CSF with low affinity (3 nM) and confers both proliferative and differentiation signals to stably transfected murine Ba/F3 cells. MATERIALS AND METHODS: To investigate whether the common beta-subunit of the GM-CSF receptor (beta c) can interact with GMER, either the entire beta-subunit or a mutant, truncated beta-subunit that completely lacks the cytoplasmic domain (beta tr) was introduced into Ba/F3 cells that express GMER, and the binding of GM-CSF as well as proliferation and differentiation responses were measured. RESULTS: Scatchard analysis showed that both GMER + beta c and GMER + beta tr bound hGM-CSF with high affinity (Kd 40 pM to 65 pM). Proliferation assays showed that the maximum growth of cells expressing GMER + beta c was identical to that of cells with GMER alone. However, proliferation of the cells that expressed GMER + beta tr was reduced by 80-95% of GMER. Dose-response curves showed that the concentration of GM-CSF required for half-maximal growth was 0.5-5.0 pM for GMER + beta c and 0.5-5 nM for GMER and GMER + beta tr. The EpoR cytoplasmic domain of GMER also undergoes ligandinducible tyrosine phosphorylation. However, the tyrosine phosphorylation did not correlate with growth in cells expressing beta tr. Coexpression of beta c with GMER in Ba/F3 cells grown in hGM-CSF markedly enhanced beta-globin mRNA expression. CONCLUSIONS: These results indicate that beta c can transduce a unique signal in association with GMER to influence both proliferative and differentiation signal pathways.

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Selected References

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  • Sieff CA, Emerson SG, Donahue RE, Nathan DG, Wang EA, Wong GG, Clark SC. Human recombinant granulocyte-macrophage colony-stimulating factor: a multilineage hematopoietin. Science. 1985 Dec 6;230(4730):1171–1173. [PubMed]
  • Martin FH, Suggs SV, Langley KE, Lu HS, Ting J, Okino KH, Morris CF, McNiece IK, Jacobsen FW, Mendiaz EA, et al. Primary structure and functional expression of rat and human stem cell factor DNAs. Cell. 1990 Oct 5;63(1):203–211. [PubMed]
  • Gearing DP, King JA, Gough NM, Nicola NA. Expression cloning of a receptor for human granulocyte-macrophage colony-stimulating factor. EMBO J. 1989 Dec 1;8(12):3667–3676. [PubMed]
  • Crosier KE, Wong GG, Mathey-Prevot B, Nathan DG, Sieff CA. A functional isoform of the human granulocyte/macrophage colony-stimulating factor receptor has an unusual cytoplasmic domain. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7744–7748. [PubMed]
  • Hayashida K, Kitamura T, Gorman DM, Arai K, Yokota T, Miyajima A. Molecular cloning of a second subunit of the receptor for human granulocyte-macrophage colony-stimulating factor (GM-CSF): reconstitution of a high-affinity GM-CSF receptor. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9655–9659. [PubMed]
  • Miyajima A, Mui AL, Ogorochi T, Sakamaki K. Receptors for granulocyte-macrophage colony-stimulating factor, interleukin-3, and interleukin-5. Blood. 1993 Oct 1;82(7):1960–1974. [PubMed]
  • Weiss M, Yokoyama C, Shikama Y, Naugle C, Druker B, Sieff CA. Human granulocyte-macrophage colony-stimulating factor receptor signal transduction requires the proximal cytoplasmic domains of the alpha and beta subunits. Blood. 1993 Dec 1;82(11):3298–3306. [PubMed]
  • Sakamaki K, Miyajima I, Kitamura T, Miyajima A. Critical cytoplasmic domains of the common beta subunit of the human GM-CSF, IL-3 and IL-5 receptors for growth signal transduction and tyrosine phosphorylation. EMBO J. 1992 Oct;11(10):3541–3549. [PubMed]
  • Jubinsky PT, Nathan DG, Wilson DJ, Sieff CA. A low-affinity human granulocyte-macrophage colony-stimulating factor/murine erythropoietin hybrid receptor functions in murine cell lines. Blood. 1993 Feb 1;81(3):587–591. [PubMed]
  • Palacios R, Steinmetz M. Il-3-dependent mouse clones that express B-220 surface antigen, contain Ig genes in germ-line configuration, and generate B lymphocytes in vivo. Cell. 1985 Jul;41(3):727–734. [PubMed]
  • Liboi E, Carroll M, D'Andrea AD, Mathey-Prevot B. Erythropoietin receptor signals both proliferation and erythroid-specific differentiation. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11351–11355. [PubMed]
  • D'Andrea AD, Yoshimura A, Youssoufian H, Zon LI, Koo JW, Lodish HF. The cytoplasmic region of the erythropoietin receptor contains nonoverlapping positive and negative growth-regulatory domains. Mol Cell Biol. 1991 Apr;11(4):1980–1987. [PMC free article] [PubMed]
  • Shikama Y, Barber DL, D'Andrea AD, Sieff CA. A constitutively activated chimeric cytokine receptor confers factor-independent growth in hematopoietic cell lines. Blood. 1996 Jul 15;88(2):455–464. [PubMed]
  • Yoshimura A, D'Andrea AD, Lodish HF. Friend spleen focus-forming virus glycoprotein gp55 interacts with the erythropoietin receptor in the endoplasmic reticulum and affects receptor metabolism. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4139–4143. [PubMed]
  • Munson PJ, Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Yoshimura A, Longmore G, Lodish HF. Point mutation in the exoplasmic domain of the erythropoietin receptor resulting in hormone-independent activation and tumorigenicity. Nature. 1990 Dec 13;348(6302):647–649. [PubMed]
  • Watowich SS, Yoshimura A, Longmore GD, Hilton DJ, Yoshimura Y, Lodish HF. Homodimerization and constitutive activation of the erythropoietin receptor. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2140–2144. [PubMed]
  • Barber DL, DeMartino JC, Showers MO, D'Andrea AD. A dominant negative erythropoietin (EPO) receptor inhibits EPO-dependent growth and blocks F-gp55-dependent transformation. Mol Cell Biol. 1994 Apr;14(4):2257–2265. [PMC free article] [PubMed]
  • Nakamura Y, Nakauchi H. A truncated erythropoietin receptor and cell death: a reanalysis. Science. 1994 Apr 22;264(5158):588–589. [PubMed]
  • Maruyama K, Miyata K, Yoshimura A. Proliferation and erythroid differentiation through the cytoplasmic domain of the erythropoietin receptor. J Biol Chem. 1994 Feb 25;269(8):5976–5980. [PubMed]
  • Yoshimura A, Lodish HF. In vitro phosphorylation of the erythropoietin receptor and an associated protein, pp130. Mol Cell Biol. 1992 Feb;12(2):706–715. [PMC free article] [PubMed]
  • Witthuhn BA, Quelle FW, Silvennoinen O, Yi T, Tang B, Miura O, Ihle JN. JAK2 associates with the erythropoietin receptor and is tyrosine phosphorylated and activated following stimulation with erythropoietin. Cell. 1993 Jul 30;74(2):227–236. [PubMed]
  • Chiba T, Nagata Y, Kishi A, Sakamaki K, Miyajima A, Yamamoto M, Engel JD, Todokoro K. Induction of erythroid-specific gene expression in lymphoid cells. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11593–11597. [PubMed]
  • Damen J, Mui AL, Hughes P, Humphries K, Krystal G. Erythropoietin-induced tyrosine phosphorylations in a high erythropoietin receptor-expressing lymphoid cell line. Blood. 1992 Oct 15;80(8):1923–1932. [PubMed]
  • Hanazono Y, Sasaki K, Nitta H, Yazaki Y, Hirai H. Erythropoietin induces tyrosine phosphorylation of the beta chain of the GM-CSF receptor. Biochem Biophys Res Commun. 1995 Mar 28;208(3):1060–1066. [PubMed]

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