Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1997 Jul 1;16(13):3866–3876. doi: 10.1093/emboj/16.13.3866

Mechanism of TGFbeta receptor inhibition by FKBP12.

Y G Chen 1, F Liu 1, J Massague 1
PMCID: PMC1170011  PMID: 9233797

Abstract

Transforming growth factor-beta (TGFbeta) signaling requires phosphorylation of the type I receptor TbetaR-I by TbetaR-II. Although TGFbeta promotes the association of TbetaR-I with TbetaR-II, these receptor components have affinity for each other which can lead to their ligand-independent activation. The immunophilin FKBP12 binds to TbetaR-I and inhibits its signaling function. We investigated the mechanism and functional significance of this effect. FKBP12 binding to TbetaR-I involves the rapamycin/Leu-Pro binding pocket of FKBP12 and a Leu-Pro sequence located next to the activating phosphorylation sites in TbetaR-I. Mutations in the binding sites of FKBP12 or TbetaR-I abolish the interaction between these proteins, leading to receptor activation in the absence of added ligand. FKBP12 does not inhibit TbetaR-I association with TbetaR-II, but inhibits TbetaR-I phosphorylation by TbetaR-II. Rapamycin, which blocks FKBP12 binding to TbetaR-I, reverses the inhibitory effect of FKBP12 on TbetaR-I phosphorylation. By impeding the activation of TGFbeta receptor complexes formed in the absence of ligand, FKBP12 may provide a safeguard against leaky signaling resulting from the innate tendency of TbetaR-I and TbetaR-II to interact with each other.

Full Text

The Full Text of this article is available as a PDF (565.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andersson S., Davis D. L., Dahlbäck H., Jörnvall H., Russell D. W. Cloning, structure, and expression of the mitochondrial cytochrome P-450 sterol 26-hydroxylase, a bile acid biosynthetic enzyme. J Biol Chem. 1989 May 15;264(14):8222–8229. [PubMed] [Google Scholar]
  2. Brillantes A. B., Ondrias K., Scott A., Kobrinsky E., Ondriasová E., Moschella M. C., Jayaraman T., Landers M., Ehrlich B. E., Marks A. R. Stabilization of calcium release channel (ryanodine receptor) function by FK506-binding protein. Cell. 1994 May 20;77(4):513–523. doi: 10.1016/0092-8674(94)90214-3. [DOI] [PubMed] [Google Scholar]
  3. Brown E. J., Albers M. W., Shin T. B., Ichikawa K., Keith C. T., Lane W. S., Schreiber S. L. A mammalian protein targeted by G1-arresting rapamycin-receptor complex. Nature. 1994 Jun 30;369(6483):756–758. doi: 10.1038/369756a0. [DOI] [PubMed] [Google Scholar]
  4. Charng M. J., Kinnunen P., Hawker J., Brand T., Schneider M. D. FKBP-12 recognition is dispensable for signal generation by type I transforming growth factor-beta receptors. J Biol Chem. 1996 Sep 20;271(38):22941–22944. doi: 10.1074/jbc.271.38.22941. [DOI] [PubMed] [Google Scholar]
  5. Chen F., Weinberg R. A. Biochemical evidence for the autophosphorylation and transphosphorylation of transforming growth factor beta receptor kinases. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1565–1569. doi: 10.1073/pnas.92.5.1565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chen R. H., Moses H. L., Maruoka E. M., Derynck R., Kawabata M. Phosphorylation-dependent interaction of the cytoplasmic domains of the type I and type II transforming growth factor-beta receptors. J Biol Chem. 1995 May 19;270(20):12235–12241. doi: 10.1074/jbc.270.20.12235. [DOI] [PubMed] [Google Scholar]
  7. Chen X., Rubock M. J., Whitman M. A transcriptional partner for MAD proteins in TGF-beta signalling. Nature. 1996 Oct 24;383(6602):691–696. doi: 10.1038/383691a0. [DOI] [PubMed] [Google Scholar]
  8. Cárcamo J., Weis F. M., Ventura F., Wieser R., Wrana J. L., Attisano L., Massagué J. Type I receptors specify growth-inhibitory and transcriptional responses to transforming growth factor beta and activin. Mol Cell Biol. 1994 Jun;14(6):3810–3821. doi: 10.1128/mcb.14.6.3810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eppert K., Scherer S. W., Ozcelik H., Pirone R., Hoodless P., Kim H., Tsui L. C., Bapat B., Gallinger S., Andrulis I. L. MADR2 maps to 18q21 and encodes a TGFbeta-regulated MAD-related protein that is functionally mutated in colorectal carcinoma. Cell. 1996 Aug 23;86(4):543–552. doi: 10.1016/s0092-8674(00)80128-2. [DOI] [PubMed] [Google Scholar]
  10. Feng X. H., Derynck R. Ligand-independent activation of transforming growth factor (TGF) beta signaling pathways by heteromeric cytoplasmic domains of TGF-beta receptors. J Biol Chem. 1996 May 31;271(22):13123–13129. doi: 10.1074/jbc.271.22.13123. [DOI] [PubMed] [Google Scholar]
  11. Feng X. H., Filvaroff E. H., Derynck R. Transforming growth factor-beta (TGF-beta)-induced down-regulation of cyclin A expression requires a functional TGF-beta receptor complex. Characterization of chimeric and truncated type I and type II receptors. J Biol Chem. 1995 Oct 13;270(41):24237–24245. doi: 10.1074/jbc.270.41.24237. [DOI] [PubMed] [Google Scholar]
  12. Franzén P., Heldin C. H., Miyazono K. The GS domain of the transforming growth factor-beta type I receptor is important in signal transduction. Biochem Biophys Res Commun. 1995 Feb 15;207(2):682–689. doi: 10.1006/bbrc.1995.1241. [DOI] [PubMed] [Google Scholar]
  13. Futer O., DeCenzo M. T., Aldape R. A., Livingston D. J. FK506 binding protein mutational analysis. Defining the surface residue contributions to stability of the calcineurin co-complex. J Biol Chem. 1995 Aug 11;270(32):18935–18940. doi: 10.1074/jbc.270.32.18935. [DOI] [PubMed] [Google Scholar]
  14. Graff J. M., Bansal A., Melton D. A. Xenopus Mad proteins transduce distinct subsets of signals for the TGF beta superfamily. Cell. 1996 May 17;85(4):479–487. doi: 10.1016/s0092-8674(00)81249-0. [DOI] [PubMed] [Google Scholar]
  15. Green J. B., Smith J. C. Graded changes in dose of a Xenopus activin A homologue elicit stepwise transitions in embryonic cell fate. Nature. 1990 Sep 27;347(6291):391–394. doi: 10.1038/347391a0. [DOI] [PubMed] [Google Scholar]
  16. Harding M. W., Galat A., Uehling D. E., Schreiber S. L. A receptor for the immunosuppressant FK506 is a cis-trans peptidyl-prolyl isomerase. Nature. 1989 Oct 26;341(6244):758–760. doi: 10.1038/341758a0. [DOI] [PubMed] [Google Scholar]
  17. Harrison R. K., Stein R. L. Substrate specificities of the peptidyl prolyl cis-trans isomerase activities of cyclophilin and FK-506 binding protein: evidence for the existence of a family of distinct enzymes. Biochemistry. 1990 Apr 24;29(16):3813–3816. doi: 10.1021/bi00468a001. [DOI] [PubMed] [Google Scholar]
  18. Holley S. A., Neul J. L., Attisano L., Wrana J. L., Sasai Y., O'Connor M. B., De Robertis E. M., Ferguson E. L. The Xenopus dorsalizing factor noggin ventralizes Drosophila embryos by preventing DPP from activating its receptor. Cell. 1996 Aug 23;86(4):607–617. doi: 10.1016/s0092-8674(00)80134-8. [DOI] [PubMed] [Google Scholar]
  19. Hoodless P. A., Haerry T., Abdollah S., Stapleton M., O'Connor M. B., Attisano L., Wrana J. L. MADR1, a MAD-related protein that functions in BMP2 signaling pathways. Cell. 1996 May 17;85(4):489–500. doi: 10.1016/s0092-8674(00)81250-7. [DOI] [PubMed] [Google Scholar]
  20. Kawabata M., Imamura T., Miyazono K., Engel M. E., Moses H. L. Interaction of the transforming growth factor-beta type I receptor with farnesyl-protein transferase-alpha. J Biol Chem. 1995 Dec 15;270(50):29628–29631. doi: 10.1074/jbc.270.50.29628. [DOI] [PubMed] [Google Scholar]
  21. Kretzschmar M., Liu F., Hata A., Doody J., Massagué J. The TGF-beta family mediator Smad1 is phosphorylated directly and activated functionally by the BMP receptor kinase. Genes Dev. 1997 Apr 15;11(8):984–995. doi: 10.1101/gad.11.8.984. [DOI] [PubMed] [Google Scholar]
  22. Lagna G., Hata A., Hemmati-Brivanlou A., Massagué J. Partnership between DPC4 and SMAD proteins in TGF-beta signalling pathways. Nature. 1996 Oct 31;383(6603):832–836. doi: 10.1038/383832a0. [DOI] [PubMed] [Google Scholar]
  23. Laiho M., Weis M. B., Massagué J. Concomitant loss of transforming growth factor (TGF)-beta receptor types I and II in TGF-beta-resistant cell mutants implicates both receptor types in signal transduction. J Biol Chem. 1990 Oct 25;265(30):18518–18524. [PubMed] [Google Scholar]
  24. Liu F., Ventura F., Doody J., Massagué J. Human type II receptor for bone morphogenic proteins (BMPs): extension of the two-kinase receptor model to the BMPs. Mol Cell Biol. 1995 Jul;15(7):3479–3486. doi: 10.1128/mcb.15.7.3479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. López-Casillas F., Wrana J. L., Massagué J. Betaglycan presents ligand to the TGF beta signaling receptor. Cell. 1993 Jul 2;73(7):1435–1444. doi: 10.1016/0092-8674(93)90368-z. [DOI] [PubMed] [Google Scholar]
  26. Macías-Silva M., Abdollah S., Hoodless P. A., Pirone R., Attisano L., Wrana J. L. MADR2 is a substrate of the TGFbeta receptor and its phosphorylation is required for nuclear accumulation and signaling. Cell. 1996 Dec 27;87(7):1215–1224. doi: 10.1016/s0092-8674(00)81817-6. [DOI] [PubMed] [Google Scholar]
  27. Massagué J. Identification of receptors for type-beta transforming growth factor. Methods Enzymol. 1987;146:174–195. doi: 10.1016/s0076-6879(87)46020-5. [DOI] [PubMed] [Google Scholar]
  28. Massagué J. TGFbeta signaling: receptors, transducers, and Mad proteins. Cell. 1996 Jun 28;85(7):947–950. doi: 10.1016/s0092-8674(00)81296-9. [DOI] [PubMed] [Google Scholar]
  29. Massagué J. The transforming growth factor-beta family. Annu Rev Cell Biol. 1990;6:597–641. doi: 10.1146/annurev.cb.06.110190.003121. [DOI] [PubMed] [Google Scholar]
  30. Moustakas A., Lin H. Y., Henis Y. I., Plamondon J., O'Connor-McCourt M. D., Lodish H. F. The transforming growth factor beta receptors types I, II, and III form hetero-oligomeric complexes in the presence of ligand. J Biol Chem. 1993 Oct 25;268(30):22215–22218. [PubMed] [Google Scholar]
  31. Okadome T., Oeda E., Saitoh M., Ichijo H., Moses H. L., Miyazono K., Kawabata M. Characterization of the interaction of FKBP12 with the transforming growth factor-beta type I receptor in vivo. J Biol Chem. 1996 Sep 6;271(36):21687–21690. doi: 10.1074/jbc.271.36.21687. [DOI] [PubMed] [Google Scholar]
  32. Piccolo S., Sasai Y., Lu B., De Robertis E. M. Dorsoventral patterning in Xenopus: inhibition of ventral signals by direct binding of chordin to BMP-4. Cell. 1996 Aug 23;86(4):589–598. doi: 10.1016/s0092-8674(00)80132-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Savage C., Das P., Finelli A. L., Townsend S. R., Sun C. Y., Baird S. E., Padgett R. W. Caenorhabditis elegans genes sma-2, sma-3, and sma-4 define a conserved family of transforming growth factor beta pathway components. Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):790–794. doi: 10.1073/pnas.93.2.790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schreiber S. L. Chemistry and biology of the immunophilins and their immunosuppressive ligands. Science. 1991 Jan 18;251(4991):283–287. doi: 10.1126/science.1702904. [DOI] [PubMed] [Google Scholar]
  35. Schreiber S. L. Immunophilin-sensitive protein phosphatase action in cell signaling pathways. Cell. 1992 Aug 7;70(3):365–368. doi: 10.1016/0092-8674(92)90158-9. [DOI] [PubMed] [Google Scholar]
  36. Sekelsky J. J., Newfeld S. J., Raftery L. A., Chartoff E. H., Gelbart W. M. Genetic characterization and cloning of mothers against dpp, a gene required for decapentaplegic function in Drosophila melanogaster. Genetics. 1995 Mar;139(3):1347–1358. doi: 10.1093/genetics/139.3.1347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Siekierka J. J., Wiederrecht G., Greulich H., Boulton D., Hung S. H., Cryan J., Hodges P. J., Sigal N. H. The cytosolic-binding protein for the immunosuppressant FK-506 is both a ubiquitous and highly conserved peptidyl-prolyl cis-trans isomerase. J Biol Chem. 1990 Dec 5;265(34):21011–21015. [PubMed] [Google Scholar]
  38. Ventura F., Doody J., Liu F., Wrana J. L., Massagué J. Reconstitution and transphosphorylation of TGF-beta receptor complexes. EMBO J. 1994 Dec 1;13(23):5581–5589. doi: 10.1002/j.1460-2075.1994.tb06895.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vivien D., Attisano L., Wrana J. L., Massagué J. Signaling activity of homologous and heterologous transforming growth factor-beta receptor kinase complexes. J Biol Chem. 1995 Mar 31;270(13):7134–7141. doi: 10.1074/jbc.270.13.7134. [DOI] [PubMed] [Google Scholar]
  40. Wang T., Danielson P. D., Li B. Y., Shah P. C., Kim S. D., Donahoe P. K. The p21(RAS) farnesyltransferase alpha subunit in TGF-beta and activin signaling. Science. 1996 Feb 23;271(5252):1120–1122. doi: 10.1126/science.271.5252.1120. [DOI] [PubMed] [Google Scholar]
  41. Wang T., Li B. Y., Danielson P. D., Shah P. C., Rockwell S., Lechleider R. J., Martin J., Manganaro T., Donahoe P. K. The immunophilin FKBP12 functions as a common inhibitor of the TGF beta family type I receptors. Cell. 1996 Aug 9;86(3):435–444. doi: 10.1016/s0092-8674(00)80116-6. [DOI] [PubMed] [Google Scholar]
  42. Weis-Garcia F., Massagué J. Complementation between kinase-defective and activation-defective TGF-beta receptors reveals a novel form of receptor cooperativity essential for signaling. EMBO J. 1996 Jan 15;15(2):276–289. [PMC free article] [PubMed] [Google Scholar]
  43. Wieser R., Wrana J. L., Massagué J. GS domain mutations that constitutively activate T beta R-I, the downstream signaling component in the TGF-beta receptor complex. EMBO J. 1995 May 15;14(10):2199–2208. doi: 10.1002/j.1460-2075.1995.tb07214.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wrana J. L., Attisano L., Cárcamo J., Zentella A., Doody J., Laiho M., Wang X. F., Massagué J. TGF beta signals through a heteromeric protein kinase receptor complex. Cell. 1992 Dec 11;71(6):1003–1014. doi: 10.1016/0092-8674(92)90395-s. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES