The Type V Transforming Growth Factor β Receptor Is the Putative Insulin-like Growth Factor-binding Protein 3 Receptor

Abstract

Insulin-like growth factor-binding protein 3 (IGFBP-3) has been shown to inhibit cell growth by IGF-dependent and -independent mechanisms. The putative cell-surface IGFBP-3 receptor that mediates the IGF-independent growth inhibition has not been identified. Here we show that recombinant human IGFBP-3 inhibits125I-transforming growth factor (TGF)-β1 binding to the type V TGF-β receptor (M r 400,000) in mink lung epithelial cells. We also demonstrate that the ∼400-kDa125I-IGFBP-3 affinity-labeled putative IGFBP-3 receptor is immunoprecipitated by specific antiserum to the type V TGF-β receptor. The 125I-IGFBP-3 affinity labeling of the putative receptor and IGFBP-3-induced growth inhibition as measured by DNA synthesis in these cells is blocked by a TGF-β1peptide antagonist. The 125I-IGFBP-3 affinity-labeled putative receptor can only be detected in cells expressing the type V TGF-β receptor, but not in cells lacking the type V TGF-β receptor. These results indicate that the type V TGF-β receptor is the putative IGFBP-3 receptor and that IGFBP-3 is a functional ligand for the type V TGF-β receptor. Insulin-like growth factor-binding protein 3 (IGFBP-3) has been shown to inhibit cell growth by IGF-dependent and -independent mechanisms. The putative cell-surface IGFBP-3 receptor that mediates the IGF-independent growth inhibition has not been identified. Here we show that recombinant human IGFBP-3 inhibits125I-transforming growth factor (TGF)-β1 binding to the type V TGF-β receptor (M r 400,000) in mink lung epithelial cells. We also demonstrate that the ∼400-kDa125I-IGFBP-3 affinity-labeled putative IGFBP-3 receptor is immunoprecipitated by specific antiserum to the type V TGF-β receptor. The 125I-IGFBP-3 affinity labeling of the putative receptor and IGFBP-3-induced growth inhibition as measured by DNA synthesis in these cells is blocked by a TGF-β1peptide antagonist. The 125I-IGFBP-3 affinity-labeled putative receptor can only be detected in cells expressing the type V TGF-β receptor, but not in cells lacking the type V TGF-β receptor. These results indicate that the type V TGF-β receptor is the putative IGFBP-3 receptor and that IGFBP-3 is a functional ligand for the type V TGF-β receptor. The type V transforming growth factor β (TGF-β) 1The abbreviations used are: TGF, transforming growth factor; IGFBP-3, insulin-like growth factor-binding protein 3; DSS, disuccinimidyl suberate; PAI-1, plasminogen activator inhibitor 1. 1The abbreviations used are: TGF, transforming growth factor; IGFBP-3, insulin-like growth factor-binding protein 3; DSS, disuccinimidyl suberate; PAI-1, plasminogen activator inhibitor 1. receptor is a 400-kDa non-proteoglycan membrane glycoprotein that co-expresses with the type I, type II, and type III TGF-β receptors in most cell types (1Massagué J. Cell. 1992; 69: 1067-1070Abstract Full Text PDF PubMed Scopus (639) Google Scholar, 2Yingling J.M. Wang X.-F. Bassing C.H. Biochim. Biophys. Acta. 1995; 1242: 115-136PubMed Google Scholar, 3O'Grady P. Kuo M.-D. Baldassare J.J. Huang S.S. Huang J.S. J. Biol. Chem. 1991; 266: 8583-8589Abstract Full Text PDF PubMed Google Scholar, 4Liu Q. Huang S.S. Huang J.S. J. Biol. Chem. 1994; 269: 9221-9226Abstract Full Text PDF PubMed Google Scholar, 5O'Grady P. Liu Q. Huang S.S. Huang J.S. J. Biol. Chem. 1992; 267: 21033-21037Abstract Full Text PDF PubMed Google Scholar). The type V TGF-β receptor as well as the type I and type II TGF-β receptors are Ser/Thr-specific protein kinases and belong to the new class of membrane receptors associated with a Ser/Thr-specific protein kinase activity (1Massagué J. Cell. 1992; 69: 1067-1070Abstract Full Text PDF PubMed Scopus (639) Google Scholar, 2Yingling J.M. Wang X.-F. Bassing C.H. Biochim. Biophys. Acta. 1995; 1242: 115-136PubMed Google Scholar, 3O'Grady P. Kuo M.-D. Baldassare J.J. Huang S.S. Huang J.S. J. Biol. Chem. 1991; 266: 8583-8589Abstract Full Text PDF PubMed Google Scholar, 4Liu Q. Huang S.S. Huang J.S. J. Biol. Chem. 1994; 269: 9221-9226Abstract Full Text PDF PubMed Google Scholar, 5O'Grady P. Liu Q. Huang S.S. Huang J.S. J. Biol. Chem. 1992; 267: 21033-21037Abstract Full Text PDF PubMed Google Scholar, 6Wrana J.L. Attisano L. Wieser R. Ventura F. Massagué J. Nature. 1994; 370: 341-346Crossref PubMed Scopus (2089) Google Scholar). The type I and type II TGF-β receptors have been shown to be important in TGF-β-induced cellular responses (1Massagué J. Cell. 1992; 69: 1067-1070Abstract Full Text PDF PubMed Scopus (639) Google Scholar, 2Yingling J.M. Wang X.-F. Bassing C.H. Biochim. Biophys. Acta. 1995; 1242: 115-136PubMed Google Scholar, 3O'Grady P. Kuo M.-D. Baldassare J.J. Huang S.S. Huang J.S. J. Biol. Chem. 1991; 266: 8583-8589Abstract Full Text PDF PubMed Google Scholar, 4Liu Q. Huang S.S. Huang J.S. J. Biol. Chem. 1994; 269: 9221-9226Abstract Full Text PDF PubMed Google Scholar, 5O'Grady P. Liu Q. Huang S.S. Huang J.S. J. Biol. Chem. 1992; 267: 21033-21037Abstract Full Text PDF PubMed Google Scholar, 6Wrana J.L. Attisano L. Wieser R. Ventura F. Massagué J. Nature. 1994; 370: 341-346Crossref PubMed Scopus (2089) Google Scholar), but the role of the type V TGF-β receptor in these responses has not been defined (3O'Grady P. Kuo M.-D. Baldassare J.J. Huang S.S. Huang J.S. J. Biol. Chem. 1991; 266: 8583-8589Abstract Full Text PDF PubMed Google Scholar, 4Liu Q. Huang S.S. Huang J.S. J. Biol. Chem. 1994; 269: 9221-9226Abstract Full Text PDF PubMed Google Scholar, 5O'Grady P. Liu Q. Huang S.S. Huang J.S. J. Biol. Chem. 1992; 267: 21033-21037Abstract Full Text PDF PubMed Google Scholar). Recently, we have demonstrated that the type V TGF-β receptor mediates TGF-β-induced growth inhibition and that both type I and type II TGF-β receptors are required for mediating maximal growth inhibition (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar). Insulin-like growth factor-binding protein 3 (IGFBP-3) is the most abundant insulin-like growth factor-binding protein in the circulation (8Baxter R.C. Martin J.L. Prog. Growth Factor Res. 1989; 1: 49-68Abstract Full Text PDF PubMed Scopus (556) Google Scholar, 9Conover C.A. J. Clin. Invest. 1991; 88: 1354-1361Crossref PubMed Scopus (82) Google Scholar, 10Clemmons D.R. Growth Regul. 1992; 2: 80-87PubMed Google Scholar, 11Martin J.L. Ballesteros M. Baxter R.C. Endocrinology. 1993; 131: 1703-1710Crossref Scopus (124) Google Scholar). In human plasma, IGFBP-3 forms an ∼140-kDa ternary complex with IGFs and an acid-labile subunit (12Jones J.I. Clemmons D.R. Endocr. Rev. 1995; 16: 3-34Crossref PubMed Google Scholar). This complex serves as a reservoir for IGFs (12Jones J.I. Clemmons D.R. Endocr. Rev. 1995; 16: 3-34Crossref PubMed Google Scholar). IGFBP-3 is produced by a variety of cell types (12Jones J.I. Clemmons D.R. Endocr. Rev. 1995; 16: 3-34Crossref PubMed Google Scholar) and appears to inhibit cell growth by IGF-dependent and -independent mechanisms (13Liu L. Delbé J. Blat C. Zapf J. Harel L. J. Cell. Physiol. 1992; 153: 15-21Crossref PubMed Scopus (53) Google Scholar, 14Oh Y. Müller H.L. Lamson G. Rosenfeld R.G. J. Biol. Chem. 1993; 268: 14964-14971Abstract Full Text PDF PubMed Google Scholar, 15Valentinis B. Bhala A. DeAngelis T. Baserga R. Cohen P. Mol. Endocrinol. 1995; 9: 361-367Crossref PubMed Google Scholar). Although several small cell membrane-associated IGFBP-3 binding proteins have recently been reported (16Delbe J. Blat C. Desauty G. Harel L. Biochem. Biophys. Res. Commun. 1991; 179: 495-501Crossref PubMed Scopus (33) Google Scholar, 17Oh Y. Müller H.L. Pham H. Rosenfeld R.G. J. Biol. Chem. 1993; 268: 26045-26048Abstract Full Text PDF PubMed Google Scholar, 18Hodgkinson S. Fowke P. Al Somai N. McQuoid M. J. Endocrinol. 1995; 145: R2-R6Crossref Scopus (19) Google Scholar), the putative IGFBP-3 receptor that mediates the IGF-independent growth inhibition has not been identified. IGFBP-3 has been implicated as a mediator of the actions of TGF-β, retinoic acid, and the tumor suppressor gene p53 (19Oh Y. Müller H.L. Ng L. Rosenfeld R.G. J. Biol. Chem. 1995; 270: 13589-13592Abstract Full Text Full Text PDF PubMed Scopus (225) Google Scholar, 20Gucev Z.S. Oh Y. Kelley K.M. Rosenfeld R.G. Cancer Res. 1996; 56: 1545-1550PubMed Google Scholar, 21Buckbinder L. Talbott R. Valasco-Miguel S. Takenaka I. Raha B. Seizinger B.R. Kley N. Nature. 1995; 377: 646-648Crossref PubMed Scopus (802) Google Scholar). Since the type V TGF-β receptor appears to play an important role in TGF-β-induced growth inhibition (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar), we tested the hypothesis that IGF-independent actions of IGFBP-3 are mediated by the type V TGF-β receptor. In this communication, we demonstrate that IGFBP-3 inhibits125I-labeled TGF-β1(125I-TGF-β1) binding to the type V TGF-β receptor in mink lung epithelial cells. We also show that125I-labeled IGFBP-3 (125I-IGFBP-3) affinity-labeled putative cell-surface IGFBP-3 receptor is immunoprecipitated by specific antiserum to the type V TGF-β receptor and that the 125I-IGFBP-3-putative receptor complex is detected only in cells expressing the type V TGF-β receptor. Finally, we show that 125I-IGFBP-3 affinity labeling of the putative IGFBP-3 receptor and IGFBP-3-induced growth inhibition can be blocked by a TGF-β1 peptide antagonist. Na125I (17 Ci/mg) and [methyl-3H]Thymidine (67 Ci/mmol) were purchased from ICN Biochemicals Inc. (Costa Mesa, CA). High molecular mass protein standards (myosin, 205 kDa; β-galactosidase, 116 kDa; phosphorylase, 97 kDa; bovine serum albumin, 66 kDa) and other chemical reagents were obtained from Sigma. Disuccinimidyl suberate (DSS) was obtained from Pierce. TGF-β1 was purchased from Austral Biologicals (San Ramon, CA). Recombinant nonglycosylated human IGFBP-3 (expressed in Escherichia coli) was provided by Celtrix Pharmaceutical Inc. (Santa Clara, CA).125I-TGF-β1 and 125I-IGFBP-3 were prepared as described previously (3O'Grady P. Kuo M.-D. Baldassare J.J. Huang S.S. Huang J.S. J. Biol. Chem. 1991; 266: 8583-8589Abstract Full Text PDF PubMed Google Scholar, 5O'Grady P. Liu Q. Huang S.S. Huang J.S. J. Biol. Chem. 1992; 267: 21033-21037Abstract Full Text PDF PubMed Google Scholar, 26O'Grady P. Huang S.S. Huang J.S. Biochem. Biophys. Res. Commun. 1991; 179: 378-385Crossref PubMed Scopus (34) Google Scholar) except 0.2 msodium phosphate buffer, pH 7.4, was used as the solvent for Sephadex G-25 column chromatography to separate 125I-IGFBP-3 from free 125I. The specific radioactivity of125I-TGF-β1 and 125I-IGFBP-3 was 1–4 × 105 cpm/ng. The antigen used to prepare specific rabbit antiserum to the type V TGF-β receptor was thyroglobulin-conjugated to a hexadecapeptide whose amino acid sequence was derived from the partial amino acid sequence of bovine type V TGF-β receptor (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar). The antiserum specifically reacted with the type V TGF-β receptor from different species, including mink, rat, mouse, cow, and human (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar). This antiserum did not react with the type I, type II, and type III TGF-β receptors on Western blot analysis and in immunoprecipitation (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar). TGF-β1 and TGF-β3peptide antagonists were synthetic pentacosapeptides whose amino acid sequences were derived from those of TGF-β1 and TGF-β3, respectively. 2S. S. Huang, Q. Liu, F. E. Johnson, Y. Konish, and J. S. Huang, submitted for publication. The IC50 values of TGF-β1 and TGF-β3 peptide antagonists for inhibiting125I-TGF-β1 (0.1 nm) binding to TGF-β receptors in mink lung epithelial cells are ∼1–2 and ∼20–30 μm, respectively.2 Human colorectal carcinoma cells transfected with neo vector only and with vector expressing type II TGF-β receptor cDNA (HCT-116 and RII-37 cells) were provided by Dr. Michael G. Brattain. (Department of Biochemistry and Molecular Biology, Medical College of Ohio, Toledo, OH)The type I and type II TGF-β receptor-defective mutant mink lung epithelial cells (R1-B and DR 26 cells) were provided by Dr. Joan Massagué (Sloan-Kettering Cancer Center, New York). Wild-type and mutant mink lung epithelial cells and other cell types were maintained in Dulbecco's modified Eagle medium containing 10% fetal calf serum. The 125I-TGF-β binding and affinity labeling were carried out as described previously (3O'Grady P. Kuo M.-D. Baldassare J.J. Huang S.S. Huang J.S. J. Biol. Chem. 1991; 266: 8583-8589Abstract Full Text PDF PubMed Google Scholar, 26O'Grady P. Huang S.S. Huang J.S. Biochem. Biophys. Res. Commun. 1991; 179: 378-385Crossref PubMed Scopus (34) Google Scholar). The specific binding of 125I-TGF-β1 was calculated by subtracting the total binding from the nonspecific binding obtained in the presence of 100-fold excess of unlabeled TGF-β1 or 10 μm TGF-β1 peptide antagonist. The125I-TGF-β1 affinity labeling of cell-surface TGF-β receptors was carried out using DSS as the cross-linking agent (3O'Grady P. Kuo M.-D. Baldassare J.J. Huang S.S. Huang J.S. J. Biol. Chem. 1991; 266: 8583-8589Abstract Full Text PDF PubMed Google Scholar, 26O'Grady P. Huang S.S. Huang J.S. Biochem. Biophys. Res. Commun. 1991; 179: 378-385Crossref PubMed Scopus (34) Google Scholar). Cells grown on 60-mm Petri dishes were incubated with 5 nm125I-IGFBP-3 (specific radioactivity: ∼1 × 105 cpm/ng) in the presence of 100-fold excess of unlabeled IGFBP-3 or inhibitors, including heparin, IGF-I, TGF-β1, and TGF-β3 peptide antagonists. After 125I-IGFBP-3 affinity labeling in the presence of DSS (3O'Grady P. Kuo M.-D. Baldassare J.J. Huang S.S. Huang J.S. J. Biol. Chem. 1991; 266: 8583-8589Abstract Full Text PDF PubMed Google Scholar, 26O'Grady P. Huang S.S. Huang J.S. Biochem. Biophys. Res. Commun. 1991; 179: 378-385Crossref PubMed Scopus (34) Google Scholar), the 125I-IGFBP-3-putative receptor complex was analyzed by 5% SDS-polyacrylamide gel electrophoresis under reducing conditions and autoradiography. After 125I-IGFBP-3 affinity labeling, the cells were detached and lysed in 100 μl of 1% Triton X-100 in 10 mm Tris-HCl, pH 7.0, 125 mm NaCl, and 1 mm EDTA. After centrifugation, the Triton X-100 extracts were then diluted 10-fold with Triton X-100-free buffer and incubated with antiserum or non-immune serum (1:100 dilution) at The were with μl of protein After with pH 7.4, Triton the were analyzed by 5% SDS-polyacrylamide gel electrophoresis under reducing conditions and autoradiography. The V TGF-β receptor complex on the was by a Cells were on dishes at and incubated with of IGFBP-3 or 10 of TGF-β1 10 peptide in Dulbecco's modified Eagle medium containing fetal calf serum. After at for the cells were with 1 of at for The cellular DNA was by a cells grown on dishes were with and 5 of IGFBP-3 or with TGF-β a for at in fetal calf serum. cellular was with to the was in gel and to membrane using 10 × The blot was at with a of and of plasminogen activator inhibitor 1 The were with × containing at TGF-β is the most growth inhibitor for epithelial cells and other cell types J. Rev. Biol. PubMed Scopus Google Scholar, A. and M. B. in Growth and Scholar, Rev. 1993; Google Scholar). have that the type V TGF-β receptor, a 400-kDa membrane glycoprotein co-expresses with the type I, type II, and type III TGF-β receptors in most cell types J.M. Wang X.-F. Bassing C.H. Biochim. Biophys. Acta. 1995; 1242: 115-136PubMed Google Scholar, 3O'Grady P. Kuo M.-D. Baldassare J.J. Huang S.S. Huang J.S. J. Biol. Chem. 1991; 266: 8583-8589Abstract Full Text PDF PubMed Google Scholar, 4Liu Q. Huang S.S. Huang J.S. J. Biol. Chem. 1994; 269: 9221-9226Abstract Full Text PDF PubMed Google Scholar, 26O'Grady P. Huang S.S. Huang J.S. Biochem. Biophys. Res. Commun. 1991; 179: 378-385Crossref PubMed Scopus (34) Google Scholar), an important role in mediating TGF-β-induced growth inhibition in mink lung epithelial cells (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar). the IGF-independent growth of IGFBP-3 is mediated by the type V TGF-β receptor or other TGF-β receptor we the of IGFBP-3 on the binding of 125I-TGF-β1 to mink lung epithelial for IGFBP-3 is also a growth shown in 1 IGFBP-3 the specific binding of a nm) or of IGFBP-3, a maximal inhibition was This partial inhibition that IGFBP-3 for binding to specific TGF-β receptor TGF-β receptor types are for IGFBP-3 we affinity labeling of cell-surface TGF-β receptors of the cells in the presence of unlabeled IGFBP-3 or 10 μm of TGF-β1peptide antagonist. The TGF-β1 peptide is a synthetic whose amino acid sequence was derived from that of shown in the type I, type II, type and type V TGF-β receptors were affinity-labeled in the presence of the cross-linking agent DSS IGFBP-3 nm) to affinity labeling of the type V TGF-β receptor, and to a the type III TGF-β receptor 1 In the the TGF-β1 peptide blocked 125I-TGF-β1 affinity labeling of TGF-β receptor types 1 These results that IGFBP-3 for binding to the type V TGF-β receptor. that IGFBP-3 to the type V TGF-β receptor with affinity or that the type V TGF-β receptor is the putative receptor for IGFBP-3, we the binding and cross-linking recombinant nonglycosylated human IGFBP-3 5 nm) to putative cell-surface receptor, by immunoprecipitation with specific antiserum to the type V TGF-β receptor (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar). 5 was to to the type V TGF-β receptor but not other TGF-β receptor shown in 125I-IGFBP-3 was to an putative receptor on the cell of mink lung epithelial cells binding and cross-linking was blocked by 100-fold excess of unlabeled IGFBP-3 or 10 peptide but not by 10 peptide and The TGF-β3peptide a whose amino acid sequence was derived from TGF-β3, has a affinity to the type V TGF-β The antiserum to the type V TGF-β receptor specifically immunoprecipitated the receptor complex 5 and and were also in the cell and in the Since the of 125I-IGFBP-3 r on SDS-polyacrylamide gel used in the was to r and 125I-IGFBP-3 has been shown to a in Y. Müller H.L. Pham H. Rosenfeld R.G. J. Biol. Chem. 1993; 268: 26045-26048Abstract Full Text PDF PubMed Google Scholar), M. Q. Liu, S. S. Huang, and J. S. Huang, be and In the 125I-IGFBP-3-putative receptor complex was in the the cells were incubated in the presence of 10 peptide to cross-linking and immunoprecipitation serum did not the 125I-IGFBP-3-putative receptor complex These results specifically to the type V TGF-β receptor in mink lung epithelial cells. the binding of 125I-IGFBP-3 to the type V TGF-β receptor, we the specific binding of to the type V TGF-β receptor in mink lung epithelial cells. shown in 125I-IGFBP-3 to the type V TGF-β receptor in these cells in a The analysis of the binding that the for 125I-IGFBP-3 binding to the type V TGF-β receptor was not Since IGFBP-3 is to IGFs with and a (8Baxter R.C. Martin J.L. Prog. Growth Factor Res. 1989; 1: 49-68Abstract Full Text PDF PubMed Scopus (556) Google Scholar, 9Conover C.A. J. Clin. Invest. 1991; 88: 1354-1361Crossref PubMed Scopus (82) Google Scholar, 10Clemmons D.R. Growth Regul. 1992; 2: 80-87PubMed Google Scholar, 11Martin J.L. Ballesteros M. Baxter R.C. Endocrinology. 1993; 131: 1703-1710Crossref Scopus (124) Google Scholar), we the of the complex and on the binding of 125I-IGFBP-3 to type V TGF-β receptor in mink lung epithelial cells. shown in at 1 of and of the 125I-IGFBP-3 specific binding to the type V TGF-β receptor was at 100 of binding to the type V TGF-β receptor The was to show the V TGF-β receptor complex in and These results that both the complex and the complex are not of binding to the type V TGF-β receptor. a TGF-β1 peptide of the 125I-IGFBP-3 binding to the type V TGF-β receptor demonstrate that the type V TGF-β receptor is the putative IGFBP-3 receptor, we the 125I-IGFBP-3 affinity labeling of putative cell-surface receptor in cells expressing and lacking the type V TGF-β receptor. shown in human colorectal carcinoma cells cells and 116 the type V TGF-β receptor (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar, J. L. L. J. J. E. S. J. Biol. Chem. 1995; 270: Full Text Full Text PDF PubMed Scopus Google Scholar), did not show the receptor complex In are to the type V TGF-β receptor P. Huang S.S. Huang J.S. Biochem. Biophys. Res. Commun. 1991; 179: 378-385Crossref PubMed Scopus (34) Google Scholar), the receptor complex 1 and The of the receptor complex in cells was blocked in the presence of 100-fold excess of unlabeled IGFBP-3 or 10 μm TGF-β1 peptide not affinity labeling of the putative cell-surface IGFBP-3 receptor in human colorectal carcinoma mink lung epithelial and cells. receptors were affinity-labeled and The 125I-IGFBP-3 affinity-labeled receptors were then analyzed by 5% SDS-polyacrylamide gel electrophoresis and autoradiography. to affinity labeling the binding of 125I-IGFBP-3 to 116 and cells was carried out in the and presence of 10 peptide not the V TGF-β receptor Since the type V TGF-β receptor has been shown to the growth in mink lung epithelial cells (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar), we the of IGFBP-3 on the of and type I and type II TGF-β receptor-defective mutant mink lung epithelial cells and M. Massagué J. J. Biol. Chem. 1991; 266: Full Text PDF PubMed Google Scholar, Massagué J. J. Biol. Chem. 1989; Full Text PDF PubMed Google Scholar, S. H. M. P. Massagué J. J. Biol. Chem. Full Text PDF PubMed Google Scholar, M. Massagué J. J. Biol. Chem. Full Text PDF PubMed Google Scholar). and cells have been shown to the type V TGF-β receptor (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar). IGFBP-3 be a specific ligand to the of the type V TGF-β receptor, not to the type I, type II, or type III TGF-β receptor with shown in IGFBP-3 or nm) a growth as measured by DNA synthesis in cells) or type II TGF-β receptor-defective mutant mink lung epithelial cells but to a in type I TGF-β receptor-defective mutant cells The growth by IGFBP-3 in these cells be blocked in the presence of TGF-β1 peptide These results indicate that IGFBP-3 a growth in cells expressing the type V TGF-β receptor. These results also the hypothesis that the type V TGF-β receptor can the growth (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar). In a P. Huang S.S. Huang J.S. Biochem. Biophys. Res. Commun. 1991; 179: 378-385Crossref PubMed Scopus (34) Google Scholar), we reported that types of carcinoma cells the type V TGF-β receptor and that cells not to TGF-β1 as measured by growth inhibition (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar). Recently, human colorectal carcinoma cells 116 cells) were shown to be in the type II TGF-β receptor J. L. L. J. J. E. S. J. Biol. Chem. 1995; 270: Full Text Full Text PDF PubMed Scopus Google Scholar). of these carcinoma cells with the type II TGF-β receptor cDNA was to the but to the growth to TGF-β J. L. L. J. J. E. S. J. Biol. Chem. 1995; 270: Full Text Full Text PDF PubMed Scopus Google Scholar). This appears to be to the of the type V TGF-β receptor in cells transfected with the neo vector only 116 cells) or with vector expressing the type II TGF-β receptor cDNA cells) (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar, J. L. L. J. J. E. S. J. Biol. Chem. 1995; 270: Full Text Full Text PDF PubMed Scopus Google Scholar). be IGFBP-3 also to inhibit growth in these 116 and RII-37 cells that not the type V TGF-β receptor not TGF-β a variety of in different cell types J. Rev. Biol. PubMed Scopus Google Scholar, A. and M. B. in Growth and Scholar, Rev. 1993; Google Scholar). In to growth the other activity of TGF-β is of and J. Rev. Biol. PubMed Scopus Google Scholar, A. and M. B. in Growth and Scholar, Rev. 1993; Google Scholar). IGFBP-3 and TGF-β we the of IGFBP-3 on the of in mink lung epithelial cells. IGFBP-3 on the of in these epithelial cells not TGF-β has been shown to be a growth a growth inhibitor for epithelial and other cell and a factor for cells J. Rev. Biol. PubMed Scopus Google Scholar, A. and M. B. in Growth and Scholar, Rev. 1993; Google Scholar). We the of IGFBP-3 on DNA synthesis in for TGF-β is a IGFBP-3 did not DNA synthesis of cells at of that IGFBP-3 is a partial of These results also the hypothesis that the type V TGF-β receptor mediates the growth in cells (7Liu, Q., Huang, S. S., and Huang, J. S. (1997) J. Biol. Chem., 272, in press.Google Scholar). IGFBP-3 has been implicated as a mediator of the actions of TGF-β, retinoic acid, and p53 (19Oh Y. Müller H.L. Ng L. Rosenfeld R.G. J. Biol. Chem. 1995; 270: 13589-13592Abstract Full Text Full Text PDF PubMed Scopus (225) Google Scholar, 20Gucev Z.S. Oh Y. Kelley K.M. Rosenfeld R.G. Cancer Res. 1996; 56: 1545-1550PubMed Google Scholar, 21Buckbinder L. Talbott R. Valasco-Miguel S. Takenaka I. Raha B. Seizinger B.R. Kley N. Nature. 1995; 377: 646-648Crossref PubMed Scopus (802) Google Scholar). to IGFBP-3 has been shown to the growth by TGF-β and retinoic acid in human carcinoma cells (19Oh Y. Müller H.L. Ng L. Rosenfeld R.G. J. Biol. Chem. 1995; 270: 13589-13592Abstract Full Text Full Text PDF PubMed Scopus (225) Google Scholar, 20Gucev Z.S. Oh Y. Kelley K.M. Rosenfeld R.G. Cancer Res. 1996; 56: 1545-1550PubMed Google Scholar, 21Buckbinder L. Talbott R. Valasco-Miguel S. Takenaka I. Raha B. Seizinger B.R. Kley N. Nature. 1995; 377: 646-648Crossref PubMed Scopus (802) Google Scholar). The functional role of IGFBP-3 in TGF-β-induced growth inhibition in other cell types is We that the IGFBP-3 by TGF-β and retinoic acid in these carcinoma cells the growth inhibition by IGFs from binding to receptor in cells and by the growth mediated by the type V TGF-β receptor in cells. is important to that ligand the type V TGF-β receptor also in the by the and of the cell receptor. Liu, J. H. S. S. Huang, S. and J. S. Huang, submitted for publication. This on also to the growth mediated by the type V TGF-β We Celtrix Pharmaceutical Inc. for recombinant nonglycosylated human Joan Massagué and Michael G. for TGF-β receptor-defective mutant mink lung epithelial cells and cells) and human colorectal carcinoma cells 116 and RII-37 S. and E. for and of the and for in the of this