RSS-Feed abonnieren
DOI: 10.1055/s-2003-41619
Subcellular Trafficking of Growth Hormone Receptor and Jak2 under Ligand Exposure
Publikationsverlauf
Received 6 January 2003
Accepted after Revision 14 April 2003
Publikationsdatum:
21. August 2003 (online)
Abstract
In CHO cells, growth hormone stimulation induces a rapid degradation of mature and precursor forms of its receptor, but does not affect Jak2 concentration. Confocal analysis of the receptor and of specific markers for subcellular localization shows that ligand exposure induced the disappearance of cell surface receptors, while some receptors seem to be sequestered in the endoplasmic reticulum (ER) and in the Golgi apparatus. Using a tagged version of Jak2 (HA-Jak2) and double immuno-fluorescence analysis with anti-HA and anti-Stat5 antibodies, we demonstrate that ligand stimulation induces Stat5 nuclear accumulation while Jak2 remains localized in the cytoplasm. Immunoblots of nuclear extracts confirm the Jak2 nuclear exclusion.
Key words
Growth hormone receptor - Jak2 - Immunolocalization - Cellular traffic
References
- 1 Isaksson O P, Eden S, Jansson J O. Mode of action of pituitary growth hormone on target cells. Annu Rev Physiol. 1985; 47 483-499
- 2 Bazan F. A novel family of growth factor receptors: a common binding domain in the growth hormone, prolactin, erythropoietin and IL-6 receptors, and p75 IL-2 receptor b-chain. Biochem Biophys Res Commun. 1989; 164 788-795
- 3 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; 255 306-312
- 4 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; 74 237-244
- 5 Finidori J, Kelly P A. Cytokine receptor signalling through two novel families of transducer molecules: Janus kinases, and signal transducers and activators of transcription. J Endocrinol. 1995; 147 11-23
- 6 van Kerkhof P, Sachse M, Klumperman J, Strous G J. Growth hormone receptor ubiquitination coincides with recruitment to clathrin-coated membrane domains. J Biol Chem. 2001; 276 3778-3784
- 7 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; 15 3806-3812
- 8 Govers R, ten B roeke, 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; 18 28-36
- 9 Moulin S, Bouzinba-Segard H, Kelly P A, Finidori J. Jak2 and proteasome activities control the availability of cell surface growth hormone receptors during ligand exposure 1. Cell Signal. 2003; 15 47-55
- 10 Moutoussamy S, Kelly P A, Finidori J. Growth hormone receptor and cytokine-receptor family signaling. Eur J Biochem. 1998; 255 1-11
- 11 Goujon L, Allevato G, Simonin G, Paquereau L, Le C am, Clark J, Nielsen J H, Djiane J, Postel-Vinay M C, Edery M, Kelly P A. Cytoplasmic domains of the growth hormone receptor necessary for signal transduction. Proc Natl Acad Sci USA. 1994; 91 957-961
- 12 Frank S J, Yi W, Zhao Y, Goldsmith J F, Gilliland G, Jiang J, Sakai I, Kraft A S. Regions of the JAK2 tyrosine kinase required for coupling to the growth hormone receptor. J Biol Chem. 1995; 270 14 776-14 785
- 13 Sorenson R L, Stout L E. Prolactin receptors and Jak2 in islets of Langerhans: an immunohistochemical analysis. Endocrinology. 1995; 136 4092-4098
- 14 Lobie P E, Ronsin B, Silvennoinen O, Haldosen L A, Norstedt G, Morel G. Constitutive nuclear localization of Janus kinases 1 and 2. Endocrinology. 1996; 137 4037-4045
- 15 Ram P A, Waxman D J. Interaction of growth hormone-activated STATs with SH2-containing phosphotyrosine phosphatase SHP-1 and nuclear JAK2 tyrosine kinase. J Biol Chem. 1997; 272 17 694-17 702
- 16 Ragimbeau J, Dondi E, Vasserot A, Romero P, Uze G, Pellegrini S. The receptor interaction region of Tyk2 contains a motif required for its nuclear localization 1. J Biol Chem. 2001; 276 30 812-30 818
- 17 Sotiropoulos A, Perrot-Applanat M, Dinerstein H, Pallier A, Postel-Vinay M C, Finidori J, Kelly P A. Distinct cytoplasmic regions of the growth hormone receptor are required for activation of Jak2 mitogen-activated protein kinase, and transcription. Endocrinology. 1994; 135 1292-1298
- 18 Sotiropoulos A, Moutoussamy S, Renaudie F, Clauss M, Kayser C, Gouilleux F, Kelly P A, Finidori J. Differential activation of Stat3 and Stat5 by distinct regions of the growth hormone receptor. Mol Endocrinol. 1996; 10 998-1009
- 19 Favre H, Benhamou A, Finidori J, Kelly P A, Edery M. Dual effects of suppressor of cytokine signaling (SOCS-2) on growth hormone signal transduction. FEBS Lett. 1999; 453 63-66
- 20 van Kerkhof P, Govers R, Alves d SC, Strous G J. Endocytosis and degradation of the growth hormone receptor are proteasome-dependent. J Biol Chem 2000 Jan. 21; 275 (3) 1575-1580
- 21 Fernandez L, Flores-Morales A, Lahuna O, Sliva D, Norstedt G, Haldosen L A, Mode A, Gustafsson J A. Desensitization of the growth hormone-induced Janus kinase 2 (Jak 2)/signal transducer and activator of transcription 5 (Stat5)-signaling pathway requires protein synthesis and phospholipase C. Endocrinology. 1998; 139 1815-1824
- 22 Alves d os, van Kerkhof P, Strous G J. The signal transduction of the growth hormone receptor is regulated by the ubiquitin/proteasome system and continues after endocytosis. J Biol Chem. 2001; 276 10 839-10 846
- 23 Lobie P E, Wood T J, Chen C M, Waters M J, Norstedt G. Nuclear translocation and anchorage of the growth hormone receptor. J Biol Chem. 1994; 269 31 735-31 746
P. A. Kelly
INSERM U584 · Faculté de Médecine Necker
156 rue de Vaugirard · 75730 Paris Cedex 15 · France
Telefon: + 33 (1) 40615318
Fax: + 33 (1) 43060443
eMail: u584@necker.fr