Abstract
Endocytosis of the growth hormone receptor (GHR) is regulated by the ubiquitin-conjugating system. A cytosolic 10 amino acid motif, referred to as the ubiquitin-dependent endocytosis (UbE) motif, is involved in the ubiquitination as well as in the endocytosis of the receptor. Proteins that are implicated in one of these processes have not been identified so far. Using a glutathione S-transferase (GST)-pulldown assay with a GST fusion protein encompassing the UbE motif of the GHR, a 35 kDa protein was purified. The protein was identified by MS as small glutamine-rich tetratricopeptide repeat (TPR)-containing protein (SGT). We found that GHR interacts with SGT. In vivo, both the precursor and the mature form of the receptor interacted with SGT. Inactivation of the ubiquitin-conjugating system did not affect the GHR-SGT interaction. Binding studies showed that the first TPR motif of SGT interacts with the UbE motif of the GHR. Taken together, these data show that SGT is a GHR-interacting protein, which binds independent of the ubiquitin-conjugating system.
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- Alele J., Jiang J., Goldsmith J. F., Yang X., Maheshwari H. G., Black R. A., Baumann G., Frank S. J. Blockade of growth hormone receptor shedding by a metalloprotease inhibitor. Endocrinology. 1998 Apr;139(4):1927–1935. doi: 10.1210/endo.139.4.5906. [DOI] [PubMed] [Google Scholar]
- Argetsinger L. S., Carter-Su C. Mechanism of signaling by growth hormone receptor. Physiol Rev. 1996 Oct;76(4):1089–1107. doi: 10.1152/physrev.1996.76.4.1089. [DOI] [PubMed] [Google Scholar]
- Baumann G. Growth hormone binding protein 2001. J Pediatr Endocrinol Metab. 2001 Apr;14(4):355–375. doi: 10.1515/jpem.2001.14.4.355. [DOI] [PubMed] [Google Scholar]
- Bazan J. F. Structural design and molecular evolution of a cytokine receptor superfamily. Proc Natl Acad Sci U S A. 1990 Sep;87(18):6934–6938. doi: 10.1073/pnas.87.18.6934. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blatch G. L., Lässle M. The tetratricopeptide repeat: a structural motif mediating protein-protein interactions. Bioessays. 1999 Nov;21(11):932–939. doi: 10.1002/(SICI)1521-1878(199911)21:11<932::AID-BIES5>3.0.CO;2-N. [DOI] [PubMed] [Google Scholar]
- Callahan M. A., Handley M. A., Lee Y. H., Talbot K. J., Harper J. W., Panganiban A. T. Functional interaction of human immunodeficiency virus type 1 Vpu and Gag with a novel member of the tetratricopeptide repeat protein family. J Virol. 1998 Oct;72(10):8461–8461. [PMC free article] [PubMed] [Google Scholar]
- Carter-Su C., Schwartz J., Smit L. S. Molecular mechanism of growth hormone action. Annu Rev Physiol. 1996;58:187–207. doi: 10.1146/annurev.ph.58.030196.001155. [DOI] [PubMed] [Google Scholar]
- Cenciarelli C., Hou D., Hsu K. C., Rellahan B. L., Wiest D. L., Smith H. T., Fried V. A., Weissman A. M. Activation-induced ubiquitination of the T cell antigen receptor. Science. 1992 Aug 7;257(5071):795–797. doi: 10.1126/science.1323144. [DOI] [PubMed] [Google Scholar]
- Cziepluch C., Kordes E., Poirey R., Grewenig A., Rommelaere J., Jauniaux J. C. Identification of a novel cellular TPR-containing protein, SGT, that interacts with the nonstructural protein NS1 of parvovirus H-1. J Virol. 1998 May;72(5):4149–4156. doi: 10.1128/jvi.72.5.4149-4156.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dupré S., Volland C., Haguenauer-Tsapis R. Membrane transport: ubiquitylation in endosomal sorting. Curr Biol. 2001 Nov 13;11(22):R932–R934. doi: 10.1016/s0960-9822(01)00558-9. [DOI] [PubMed] [Google Scholar]
- Fonte Virginia, Kapulkin Wadim Jan, Kapulkin Vadim, Taft Andrew, Fluet Amy, Friedman David, Link Christopher D. Interaction of intracellular beta amyloid peptide with chaperone proteins. Proc Natl Acad Sci U S A. 2002 Jun 27;99(14):9439–9444. doi: 10.1073/pnas.152313999. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Galcheva-Gargova Z., Theroux S. J., Davis R. J. The epidermal growth factor receptor is covalently linked to ubiquitin. Oncogene. 1995 Dec 21;11(12):2649–2655. [PubMed] [Google Scholar]
- Gent Jürgen, van Kerkhof Peter, Roza Marcel, Bu Guojun, Strous Ger J. Ligand-independent growth hormone receptor dimerization occurs in the endoplasmic reticulum and is required for ubiquitin system-dependent endocytosis. Proc Natl Acad Sci U S A. 2002 Jul 8;99(15):9858–9863. doi: 10.1073/pnas.152294299. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Govers R., ten Broeke T., van Kerkhof P., Schwartz A. L., Strous G. J. Identification of a novel ubiquitin conjugation motif, required for ligand-induced internalization of the growth hormone receptor. EMBO J. 1999 Jan 4;18(1):28–36. doi: 10.1093/emboj/18.1.28. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Govers R., van Kerkhof P., Schwartz A. L., Strous G. J. Linkage of the ubiquitin-conjugating system and the endocytic pathway in ligand-induced internalization of the growth hormone receptor. EMBO J. 1997 Aug 15;16(16):4851–4858. doi: 10.1093/emboj/16.16.4851. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hammond D. E., Urbé S., Vande Woude G. F., Clague M. J. Down-regulation of MET, the receptor for hepatocyte growth factor. Oncogene. 2001 May 17;20(22):2761–2770. doi: 10.1038/sj.onc.1204475. [DOI] [PubMed] [Google Scholar]
- Herrington J., Carter-Su C. Signaling pathways activated by the growth hormone receptor. Trends Endocrinol Metab. 2001 Aug;12(6):252–257. doi: 10.1016/s1043-2760(01)00423-4. [DOI] [PubMed] [Google Scholar]
- Hershko A., Ciechanover A. The ubiquitin system for protein degradation. Annu Rev Biochem. 1992;61:761–807. doi: 10.1146/annurev.bi.61.070192.003553. [DOI] [PubMed] [Google Scholar]
- Ihle J. N., Kerr I. M. Jaks and Stats in signaling by the cytokine receptor superfamily. Trends Genet. 1995 Feb;11(2):69–74. doi: 10.1016/s0168-9525(00)89000-9. [DOI] [PubMed] [Google Scholar]
- Imai Yuzuru, Soda Mariko, Hatakeyama Shigetsugu, Akagi Takumi, Hashikawa Tsutomu, Nakayama Kei Ichi, Takahashi Ryosuke. CHIP is associated with Parkin, a gene responsible for familial Parkinson's disease, and enhances its ubiquitin ligase activity. Mol Cell. 2002 Jul;10(1):55–67. doi: 10.1016/s1097-2765(02)00583-x. [DOI] [PubMed] [Google Scholar]
- Jeffers M., Taylor G. A., Weidner K. M., Omura S., Vande Woude G. F. Degradation of the Met tyrosine kinase receptor by the ubiquitin-proteasome pathway. Mol Cell Biol. 1997 Feb;17(2):799–808. doi: 10.1128/mcb.17.2.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jiang J., Ballinger C. A., Wu Y., Dai Q., Cyr D. M., Höhfeld J., Patterson C. CHIP is a U-box-dependent E3 ubiquitin ligase: identification of Hsc70 as a target for ubiquitylation. J Biol Chem. 2001 Sep 13;276(46):42938–42944. doi: 10.1074/jbc.M101968200. [DOI] [PubMed] [Google Scholar]
- Kitagawa K., Skowyra D., Elledge S. J., Harper J. W., Hieter P. SGT1 encodes an essential component of the yeast kinetochore assembly pathway and a novel subunit of the SCF ubiquitin ligase complex. Mol Cell. 1999 Jul;4(1):21–33. doi: 10.1016/s1097-2765(00)80184-7. [DOI] [PubMed] [Google Scholar]
- Levkowitz G., Waterman H., Zamir E., Kam Z., Oved S., Langdon W. Y., Beguinot L., Geiger B., Yarden Y. c-Cbl/Sli-1 regulates endocytic sorting and ubiquitination of the epidermal growth factor receptor. Genes Dev. 1998 Dec 1;12(23):3663–3674. doi: 10.1101/gad.12.23.3663. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu F. H., Wu S. J., Hu S. M., Hsiao C. D., Wang C. Specific interaction of the 70-kDa heat shock cognate protein with the tetratricopeptide repeats. J Biol Chem. 1999 Nov 26;274(48):34425–34432. doi: 10.1074/jbc.274.48.34425. [DOI] [PubMed] [Google Scholar]
- Miyata Y., Chambraud B., Radanyi C., Leclerc J., Lebeau M. C., Renoir J. M., Shirai R., Catelli M. G., Yahara I., Baulieu E. E. Phosphorylation of the immunosuppressant FK506-binding protein FKBP52 by casein kinase II: regulation of HSP90-binding activity of FKBP52. Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14500–14505. doi: 10.1073/pnas.94.26.14500. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miyazawa K., Toyama K., Gotoh A., Hendrie P. C., Mantel C., Broxmeyer H. E. Ligand-dependent polyubiquitination of c-kit gene product: a possible mechanism of receptor down modulation in M07e cells. Blood. 1994 Jan 1;83(1):137–145. [PubMed] [Google Scholar]
- Mori S., Kanaki H., Tanaka K., Morisaki N., Saito Y. Ligand-activated platelet-derived growth factor beta-receptor is degraded through proteasome-dependent proteolytic pathway. Biochem Biophys Res Commun. 1995 Dec 5;217(1):224–229. doi: 10.1006/bbrc.1995.2767. [DOI] [PubMed] [Google Scholar]
- Scheufler C., Brinker A., Bourenkov G., Pegoraro S., Moroder L., Bartunik H., Hartl F. U., Moarefi I. Structure of TPR domain-peptide complexes: critical elements in the assembly of the Hsp70-Hsp90 multichaperone machine. Cell. 2000 Apr 14;101(2):199–210. doi: 10.1016/S0092-8674(00)80830-2. [DOI] [PubMed] [Google Scholar]
- Shevchenko A., Wilm M., Vorm O., Mann M. Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem. 1996 Mar 1;68(5):850–858. doi: 10.1021/ac950914h. [DOI] [PubMed] [Google Scholar]
- Small I. D., Peeters N. The PPR motif - a TPR-related motif prevalent in plant organellar proteins. Trends Biochem Sci. 2000 Feb;25(2):46–47. doi: 10.1016/s0968-0004(99)01520-0. [DOI] [PubMed] [Google Scholar]
- Strous G. J., Govers R. The ubiquitin-proteasome system and endocytosis. J Cell Sci. 1999 May;112(Pt 10):1417–1423. doi: 10.1242/jcs.112.10.1417. [DOI] [PubMed] [Google Scholar]
- Strous G. J., van Kerkhof P., Govers R., Ciechanover A., Schwartz A. L. The ubiquitin conjugation system is required for ligand-induced endocytosis and degradation of the growth hormone receptor. EMBO J. 1996 Aug 1;15(15):3806–3812. [PMC free article] [PubMed] [Google Scholar]
- Tobaben S., Thakur P., Fernández-Chacón R., Südhof T. C., Rettig J., Stahl B. A trimeric protein complex functions as a synaptic chaperone machine. Neuron. 2001 Sep 27;31(6):987–999. doi: 10.1016/s0896-6273(01)00427-5. [DOI] [PubMed] [Google Scholar]
- Tsuchiya Naoto, Fukuda Hirokazu, Sugimura Takashi, Nagao Minako, Nakagama Hitoshi. LRP130, a protein containing nine pentatricopeptide repeat motifs, interacts with a single-stranded cytosine-rich sequence of mouse hypervariable minisatellite Pc-1. Eur J Biochem. 2002 Jun;269(12):2927–2933. doi: 10.1046/j.1432-1033.2002.02966.x. [DOI] [PubMed] [Google Scholar]
- Waters M. J., Friesen H. G. Purification and partial characterization of a nonprimate growth hormone receptor. J Biol Chem. 1979 Jul 25;254(14):6815–6825. [PubMed] [Google Scholar]
- Wu S. J., Liu F. H., Hu S. M., Wang C. Different combinations of the heat-shock cognate protein 70 (hsc70) C-terminal functional groups are utilized to interact with distinct tetratricopeptide repeat-containing proteins. Biochem J. 2001 Oct 15;359(Pt 2):419–426. doi: 10.1042/0264-6021:3590419. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhang Y., Jiang J., Black R. A., Baumann G., Frank S. J. Tumor necrosis factor-alpha converting enzyme (TACE) is a growth hormone binding protein (GHBP) sheddase: the metalloprotease TACE/ADAM-17 is critical for (PMA-induced) GH receptor proteolysis and GHBP generation. Endocrinology. 2000 Dec;141(12):4342–4348. doi: 10.1210/endo.141.12.7858. [DOI] [PubMed] [Google Scholar]
- de Vos A. M., Ultsch M., Kossiakoff A. A. Human growth hormone and extracellular domain of its receptor: crystal structure of the complex. Science. 1992 Jan 17;255(5042):306–312. doi: 10.1126/science.1549776. [DOI] [PubMed] [Google Scholar]
- van Kerkhof P., Alves dos Santos C. M., Sachse M., Klumperman J., Bu G., Strous G. J. Proteasome inhibitors block a late step in lysosomal transport of selected membrane but not soluble proteins. Mol Biol Cell. 2001 Aug;12(8):2556–2566. doi: 10.1091/mbc.12.8.2556. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Kerkhof P., Strous G. J. The ubiquitin-proteasome pathway regulates lysosomal degradation of the growth hormone receptor and its ligand. Biochem Soc Trans. 2001 Aug;29(Pt 4):488–493. doi: 10.1042/bst0290488. [DOI] [PubMed] [Google Scholar]