Abstract
TGF-β is a prototype of the TGF-β cytokine superfamily and exerts multiple regulatory effects on cell activities. It signals through two types of membrane-bound serine/threonine kinase receptors. Upon TGF-β binding, the type II receptor TβRII recruits the type I receptor TβRI and form a functional heterocomplex. TβRII trans-phosphorylates the GS region of TβRI, thus triggering its kinase activity. Activated TβRI proceeds to activate downstream Smad2/3. Signal intensity and duration through the availability, activity and destiny of TGF-β receptors are finely controlled by multiple posttranslational modifications such as phosphorylation, ubiquitination, and neddylation. This chapter introduces methods for examination of these modifications of TGF-β receptors.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Moustakas A, Heldin CH (2009) The regulation of TGFbeta signal transduction. Development 136:3699–3714
Massague J (2012) TGFbeta signalling in context. Nat Rev Mol Cell Biol 13:616–630
Ikushima H, Miyazono K (2012) TGF-beta signal transduction spreading to a wider field: a broad variety of mechanisms for context-dependent effects of TGF-beta. Cell Tissue Res 347:37–49
Schmierer B, Hill CS (2007) TGFbeta-SMAD signal transduction: molecular specificity and functional flexibility. Nat Rev Mol Cell Biol 8:970–982
Feng XH, Derynck R (2005) Specificity and versatility in TGF-beta signaling through Smads. Annu Rev Cell Dev Biol 21:659–693
Derynck R, Miyazono K (2008) The TGF-beta family. Cold Spring Harbor Laboratory Press, New York
Derynck R, Feng XH (1997) TGF-beta receptor signaling. Biochim Biophys Acta 1333:F105–F150
Massague J (1998) TGF-beta signal transduction. Annu Rev Biochem 67:753–791
Massague J, Chen YG (2000) Controlling TGF-beta signaling. Genes Dev 14:627–644
ten Dijke P, Hill CS (2004) New insights into TGF-beta-Smad signalling. Trends Biochem Sci 29:265–273
Heldin CH, Miyazono K, ten Dijke P (1997) TGF-beta signalling from cell membrane to nucleus through SMAD proteins. Nature 390:465–471
Huang F, Chen YG (2012) Regulation of TGF-beta receptor activity. Cell Biosci 2:9
Xu P, Liu J, Derynck R (2012) Post-translational regulation of TGF-beta receptor and Smad signaling. FEBS Lett 586:1871–1884
Wrighton KH, Lin X, Feng XH (2009) Phospho-control of TGF-beta superfamily signaling. Cell Res 19:8–20
Yan X, Chen YG (2011) Smad7: not only a regulator, but also a cross-talk mediator of TGF-beta signalling. Biochem J 434:1–10
Kang JS, Saunier EF, Akhurst RJ, Derynck R (2008) The type I TGF-beta receptor is covalently modified and regulated by sumoylation. Nat Cell Biol 10:654–664
Lee MK, Pardoux C, Hall MC, Lee PS, Warburton D, Qing J, Smith SM, Derynck R (2007) TGF-beta activates Erk MAP kinase signalling through direct phosphorylation of ShcA. EMBO J 26:3957–3967
Ehrlich M, Horbelt D, Marom B, Knaus P, Henis YI (2011) Homomeric and heteromeric complexes among TGF-beta and BMP receptors and their roles in signaling. Cell Signal 23:1424–1432
Zhang W, Jiang Y, Wang Q, Ma X, Xiao Z, Zuo W, Fang X, Chen YG (2009) Single-molecule imaging reveals transforming growth factor-beta-induced type II receptor dimerization. Proc Natl Acad Sci U S A 106:15679–15683
Zhang W, Yuan J, Yang Y, Xu L, Wang Q, Zuo W, Fang X, Chen YG (2010) Monomeric type I and type III transforming growth factor-beta receptors and their dimerization revealed by single-molecule imaging. Cell Res 20:1216–1223
Huang T, David L, Mendoza V, Yang Y, Villarreal M, De K, Sun L, Fang X, Lopez-Casillas F, Wrana JL, Hinck AP (2011) TGF-beta signalling is mediated by two autonomously functioning TbetaRI:TbetaRII pairs. EMBO J 30:1263–1276
Huse M, Chen YG, Massague J, Kuriyan J (1999) Crystal structure of the cytoplasmic domain of the type I TGF beta receptor in complex with FKBP12. Cell 96:425–436
Chen YG, Liu F, Massague J (1997) Mechanism of TGFbeta receptor inhibition by FKBP12. EMBO J 16:3866–3876
Souchelnytskyi S, ten Dijke P, Miyazono K, Heldin CH (1996) Phosphorylation of Ser165 in TGF-beta type I receptor modulates TGF-beta1-induced cellular responses. EMBO J 15:6231–6240
Bennett D, Alphey L (2002) PP1 binds Sara and negatively regulates Dpp signaling in Drosophila melanogaster. Nat Genet 31:419–423
Shi W, Sun C, He B, Xiong W, Shi X, Yao D, Cao X (2004) GADD34-PP1c recruited by Smad7 dephosphorylates TGFbeta type I receptor. J Cell Biol 164:291–300
Valdimarsdottir G, Goumans MJ, Itoh F, Itoh S, Heldin CH, ten Dijke P (2006) Smad7 and protein phosphatase 1alpha are critical determinants in the duration of TGF-beta/ALK1 signaling in endothelial cells. BMC Cell Biol 7:16
Batut J, Schmierer B, Cao J, Raftery LA, Hill CS, Howell M (2008) Two highly related regulatory subunits of PP2A exert opposite effects on TGF-beta/Activin/Nodal signalling. Development 135:2927–2937
Griswold-Prenner I, Kamibayashi C, Maruoka EM, Mumby MC, Derynck R (1998) Physical and functional interactions between type I transforming growth factor beta receptors and Balpha, a WD-40 repeat subunit of phosphatase 2A. Mol Cell Biol 18:6595–6604
Varshavsky A (2012) The ubiquitin system, an immense realm. Annu Rev Biochem 81:167–176
Soond SM, Chantry A (2011) How ubiquitination regulates the TGF-beta signalling pathway: new insights and new players: new isoforms of ubiquitin-activating enzymes in the E1-E3 families join the game. Bioessays 33:749–758
Inoue Y, Imamura T (2008) Regulation of TGF-beta family signaling by E3 ubiquitin ligases. Cancer Sci 99:2107–2112
Isasa M, Zuin A, Crosas B (2012) Integration of multiple ubiquitin signals in proteasome regulation. Methods Mol Biol 910:337–370
Kang JS, Liu C, Derynck R (2009) New regulatory mechanisms of TGF-beta receptor function. Trends Cell Biol 19:385–394
Zhang L, Zhou F, Drabsch Y, Gao R, Snaar-Jagalska BE, Mickanin C, Huang H, Sheppard KA, Porter JA, Lu CX, Ten Dijke P (2012) USP4 is regulated by AKT phosphorylation and directly deubiquitylates TGF-beta type I receptor. Nat Cell Biol 14:717–726
Al-Salihi MA, Herhaus L, Macartney T, Sapkota GP (2012) USP11 augments TGFbeta signalling by deubiquitylating ALK5. Open Biol 2:120063
Eichhorn PJ, Rodon L, Gonzalez-Junca A, Dirac A, Gili M, Martinez-Saez E, Aura C, Barba I, Peg V, Prat A, Cuartas I, Jimenez J, Garcia-Dorado D, Sahuquillo J, Bernards R, Baselga J, Seoane J (2012) USP15 stabilizes TGF-beta receptor I and promotes oncogenesis through the activation of TGF-beta signaling in glioblastoma. Nat Med 18:429–435
Wicks SJ, Haros K, Maillard M, Song L, Cohen RE, Dijke PT, Chantry A (2005) The deubiquitinating enzyme UCH37 interacts with Smads and regulates TGF-beta signalling. Oncogene 24:8080–8084
Yan X, Zhang J, Pan L, Wang P, Xue H, Zhang L, Gao X, Zhao X, Ning Y, Chen YG (2011) TSC-22 promotes transforming growth factor beta-mediated cardiac myofibroblast differentiation by antagonizing Smad7 activity. Mol Cell Biol 31:3700–3709
Xu L (2011) Averting a roadblock in transforming growth factor beta signaling. Mol Cell Biol 31:3684–3686
Watson IR, Irwin MS, Ohh M (2011) NEDD8 pathways in cancer. Sine Quibus Non Cancer Cell 19:168–176
Zuo W, Huang F, Chiang YJ, Li M, Du J, Ding Y, Zhang T, Lee HW, Jeong LS, Chen Y, Deng H, Feng XH, Luo S, Gao C, Chen YG (2013) c-Cbl-mediated neddylation antagonizes ubiquitination and degradation of the TGF-beta type II receptor. Mol Cell 49:499–510
Le Roy C, Wrana JL (2005) Clathrin- and non-clathrin-mediated endocytic regulation of cell signalling. Nat Rev Mol Cell Biol 6:112–126
Chen YG (2009) Endocytic regulation of TGF-beta signaling. Cell Res 19:58–70
Mu Y, Sundar R, Thakur N, Ekman M, Gudey SK, Yakymovych M, Hermansson A, Dimitriou H, Bengoechea-Alonso MT, Ericsson J, Heldin CH, Landstrom M (2011) TRAF6 ubiquitinates TGFbeta type I receptor to promote its cleavage and nuclear translocation in cancer. Nat Commun 2:330
Chandra M, Zang S, Li H, Zimmerman LJ, Champer J, Tsuyada A, Chow A, Zhou W, Yu Y, Gao H, Ren X, Lin RJ, Wang SE (2012) Nuclear translocation of type I transforming growth factor beta receptor confers a novel function in RNA processing. Mol Cell Biol 32:2183–2195
Liu C, Xu P, Lamouille S, Xu J, Derynck R (2009) TACE-mediated ectodomain shedding of the type I TGF-beta receptor downregulates TGF-beta signaling. Mol Cell 35:26–36
Wang X, Inoue S, Gu J, Miyoshi E, Noda K, Li W, Mizuno-Horikawa Y, Nakano M, Asahi M, Takahashi M, Uozumi N, Ihara S, Lee SH, Ikeda Y, Yamaguchi Y, Aze Y, Tomiyama Y, Fujii J, Suzuki K, Kondo A, Shapiro SD, Lopez-Otin C, Kuwaki T, Okabe M, Honke K, Taniguchi N (2005) Dysregulation of TGF-beta1 receptor activation leads to abnormal lung development and emphysema-like phenotype in core fucose-deficient mice. Proc Natl Acad Sci U S A 102:15791–15796
Partridge EA, Le Roy C, Di Guglielmo GM, Pawling J, Cheung P, Granovsky M, Nabi IR, Wrana JL, Dennis JW (2004) Regulation of cytokine receptors by Golgi N-glycan processing and endocytosis. Science 306:120–124
Kim YW, Park J, Lee HJ, Lee SY, Kim SJ (2012) TGF-beta sensitivity is determined by N-linked glycosylation of the type II TGF-beta receptor. Biochem J 445:403–411
Lin H, Wang D, Wu T, Dong C, Shen N, Sun Y, Sun Y, Xie H, Wang N, Shan L (2011) Blocking core fucosylation of TGF-beta1 receptors downregulates their functions and attenuates the epithelial-mesenchymal transition of renal tubular cells. Am J Physiol Renal Physiol 300:F1017–F1025
Boyd FT, Massague J (1989) Transforming growth factor-beta inhibition of epithelial cell proliferation linked to the expression of a 53-kDa membrane receptor. J Biol Chem 264:2272–2278
Wrana JL, Attisano L, Carcamo J, Zentella A, Doody J, Laiho M, Wang XF, Massague J (1992) TGF beta signals through a heteromeric protein kinase receptor complex. Cell 71:1003–1014
Lonn P, Moren A, Raja E, Dahl M, Moustakas A (2009) Regulating the stability of TGFbeta receptors and Smads. Cell Res 19:21–35
Acknowledgement
We thank Dr. Fei Huang for critical reading of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Yan, X., Chen, YG. (2016). Posttranslational Modifications of TGF-β Receptors. In: Feng, XH., Xu, P., Lin, X. (eds) TGF-β Signaling. Methods in Molecular Biology, vol 1344. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2966-5_3
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2966-5_3
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2965-8
Online ISBN: 978-1-4939-2966-5
eBook Packages: Springer Protocols