Abstract
The major function of the ubiquitin-conjugating system is the targeting of cytosolic and nuclear proteins for degradation by the proteasome. Recently, ubiquitin conjugation has been implicated in the downregulation of signalling receptors such as the mammalian growth hormone receptor (GHR) and the alpha-factor receptor in yeast. By examining truncated receptors, the internalization-deficient receptor mutant F327A and conditions under which clathrin-mediated GHR endocytosis is inhibited, we show here that GHR ubiquitination and ligand-induced GHR internalization are coupled events. Previously, we had shown that GHR endocytosis is dependent on an intact ubiquitination system. Here we present evidence that GHR ubiquitination depends on an intact endocytic pathway. Our data indicate that the ubiquitin-conjugating system and the endocytic pathway interact at the cytoplasmic tail of the GHR at the plasma membrane, where they cooperate to regulate internalization of the GHR.
Full Text
The Full Text of this article is available as a PDF (439.5 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Argetsinger L. S., Campbell G. S., Yang X., Witthuhn B. A., Silvennoinen O., Ihle J. N., Carter-Su C. Identification of JAK2 as a growth hormone receptor-associated tyrosine kinase. Cell. 1993 Jul 30;74(2):237–244. doi: 10.1016/0092-8674(93)90415-m. [DOI] [PubMed] [Google Scholar]
- Chamness G. C., McGuire W. L. Scatchard plots: common errors in correction and interpretation. Steroids. 1975 Oct;26(4):538–542. doi: 10.1016/0039-128x(75)90073-2. [DOI] [PubMed] [Google Scholar]
- Cunningham B. C., Ultsch M., De Vos A. M., Mulkerrin M. G., Clauser K. R., Wells J. A. Dimerization of the extracellular domain of the human growth hormone receptor by a single hormone molecule. Science. 1991 Nov 8;254(5033):821–825. doi: 10.1126/science.1948064. [DOI] [PubMed] [Google Scholar]
- Dittrich E., Haft C. R., Muys L., Heinrich P. C., Graeve L. A di-leucine motif and an upstream serine in the interleukin-6 (IL-6) signal transducer gp130 mediate ligand-induced endocytosis and down-regulation of the IL-6 receptor. J Biol Chem. 1996 Mar 8;271(10):5487–5494. doi: 10.1074/jbc.271.10.5487. [DOI] [PubMed] [Google Scholar]
- Garippa R. J., Johnson A., Park J., Petrush R. L., McGraw T. E. The carboxyl terminus of GLUT4 contains a serine-leucine-leucine sequence that functions as a potent internalization motif in Chinese hamster ovary cells. J Biol Chem. 1996 Aug 23;271(34):20660–20668. doi: 10.1074/jbc.271.34.20660. [DOI] [PubMed] [Google Scholar]
- Gorman C. M., Howard B. H., Reeves R. Expression of recombinant plasmids in mammalian cells is enhanced by sodium butyrate. Nucleic Acids Res. 1983 Nov 11;11(21):7631–7648. doi: 10.1093/nar/11.21.7631. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haft C. R., Klausner R. D., Taylor S. I. Involvement of dileucine motifs in the internalization and degradation of the insulin receptor. J Biol Chem. 1994 Oct 21;269(42):26286–26294. [PubMed] [Google Scholar]
- Hicke L., Riezman H. Ubiquitination of a yeast plasma membrane receptor signals its ligand-stimulated endocytosis. Cell. 1996 Jan 26;84(2):277–287. doi: 10.1016/s0092-8674(00)80982-4. [DOI] [PubMed] [Google Scholar]
- Hiller M. M., Finger A., Schweiger M., Wolf D. H. ER degradation of a misfolded luminal protein by the cytosolic ubiquitin-proteasome pathway. Science. 1996 Sep 20;273(5282):1725–1728. doi: 10.1126/science.273.5282.1725. [DOI] [PubMed] [Google Scholar]
- Ihle J. N. Cytokine receptor signalling. Nature. 1995 Oct 19;377(6550):591–594. doi: 10.1038/377591a0. [DOI] [PubMed] [Google Scholar]
- Ilondo M. M., Courtoy P. J., Geiger D., Carpentier J. L., Rousseau G. G., De Meyts P. Intracellular potassium depletion in IM-9 lymphocytes suppresses the slowly dissociating component of human growth hormone binding and the down-regulation of its receptors but does not affect insulin receptors. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6460–6464. doi: 10.1073/pnas.83.17.6460. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ilondo M. M., Smal J., de Meyts P., Courtoy P. J. Comparison of the effects of hypertonic sucrose and intracellular potassium depletion on growth hormone receptor binding kinetics and down-regulation in IM-9 cells: evidence for a sequential block of receptor--mediated endocytosis. Endocrinology. 1991 Mar;128(3):1597–1602. doi: 10.1210/endo-128-3-1597. [DOI] [PubMed] [Google Scholar]
- Jennissen H. P. Ubiquitin and the enigma of intracellular protein degradation. Eur J Biochem. 1995 Jul 1;231(1):1–30. [PubMed] [Google Scholar]
- Jentsch S. The ubiquitin-conjugation system. Annu Rev Genet. 1992;26:179–207. doi: 10.1146/annurev.ge.26.120192.001143. [DOI] [PubMed] [Google Scholar]
- Lee H. W., Smith L., Pettit G. R., Vinitsky A., Smith J. B. Ubiquitination of protein kinase C-alpha and degradation by the proteasome. J Biol Chem. 1996 Aug 30;271(35):20973–20976. [PubMed] [Google Scholar]
- Leung D. W., Spencer S. A., Cachianes G., Hammonds R. G., Collins C., Henzel W. J., Barnard R., Waters M. J., Wood W. I. Growth hormone receptor and serum binding protein: purification, cloning and expression. Nature. 1987 Dec 10;330(6148):537–543. doi: 10.1038/330537a0. [DOI] [PubMed] [Google Scholar]
- Li C. C., Dai R. M., Longo D. L. Inactivation of NF-kappa B inhibitor I kappa B alpha: ubiquitin-dependent proteolysis and its degradation product. Biochem Biophys Res Commun. 1995 Oct 4;215(1):292–301. doi: 10.1006/bbrc.1995.2465. [DOI] [PubMed] [Google Scholar]
- Murphy L. J., Lazarus L. The mouse fibroblast growth hormone receptor: ligand processing and receptor modulation and turnover. Endocrinology. 1984 Oct;115(4):1625–1632. doi: 10.1210/endo-115-4-1625. [DOI] [PubMed] [Google Scholar]
- Qu D., Teckman J. H., Omura S., Perlmutter D. H. Degradation of a mutant secretory protein, alpha1-antitrypsin Z, in the endoplasmic reticulum requires proteasome activity. J Biol Chem. 1996 Sep 13;271(37):22791–22795. doi: 10.1074/jbc.271.37.22791. [DOI] [PubMed] [Google Scholar]
- Scheffner M., Werness B. A., Huibregtse J. M., Levine A. J., Howley P. M. The E6 oncoprotein encoded by human papillomavirus types 16 and 18 promotes the degradation of p53. Cell. 1990 Dec 21;63(6):1129–1136. doi: 10.1016/0092-8674(90)90409-8. [DOI] [PubMed] [Google Scholar]
- Seufert W., Futcher B., Jentsch S. Role of a ubiquitin-conjugating enzyme in degradation of S- and M-phase cyclins. Nature. 1995 Jan 5;373(6509):78–81. doi: 10.1038/373078a0. [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]
- Strous G. J., van Kerkhof P., Govers R., Rotwein P., Schwartz A. L. Growth hormone-induced signal tranduction depends on an intact ubiquitin system. J Biol Chem. 1997 Jan 3;272(1):40–43. doi: 10.1074/jbc.272.1.40. [DOI] [PubMed] [Google Scholar]
- Sudakin V., Ganoth D., Dahan A., Heller H., Hershko J., Luca F. C., Ruderman J. V., Hershko A. The cyclosome, a large complex containing cyclin-selective ubiquitin ligase activity, targets cyclins for destruction at the end of mitosis. Mol Biol Cell. 1995 Feb;6(2):185–197. doi: 10.1091/mbc.6.2.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Winston L. A., Bertics P. J. Growth hormone stimulates the tyrosine phosphorylation of 42- and 45-kDa ERK-related proteins. J Biol Chem. 1992 Mar 5;267(7):4747–4751. [PubMed] [Google Scholar]
- Yarden Y., Escobedo J. A., Kuang W. J., Yang-Feng T. L., Daniel T. O., Tremble P. M., Chen E. Y., Ando M. E., Harkins R. N., Francke U. Structure of the receptor for platelet-derived growth factor helps define a family of closely related growth factor receptors. Nature. 1986 Sep 18;323(6085):226–232. doi: 10.1038/323226a0. [DOI] [PubMed] [Google Scholar]