Endocrinology, Vol 132, 101-108, Copyright © 1993 by Endocrine Society

ARTICLES

Stimulation of tyrosine phosphorylation in human cells by activation of the growth hormone receptor

CM Silva, MJ Weber and MO Thorner
Department of Medicine, University of Virginia Health Sciences Center, Charlottesville 22908.

To investigate the possibility that tyrosine phosphorylation of cellular proteins might play a role in GH receptor signaling, we have studied tyrosine phosphorylation induced by GH and by GH mutants in the human lymphocyte line IM-9, a homologous cell system which is known to respond to GH by increased proliferation. IM-9 cells were treated with physiological concentrations of recombinant human GH (rhGH). Protein lysates from these cells were then analyzed by polyacrylamide gel electrophoresis, transferred to nitrocellulose, and probed with an antibody specific for phosphotyrosine. rhGH stimulated the tyrosine phosphorylation of two proteins having M(r) of approximately 93,000 and 120,000. Tyrosine phosphorylation of these proteins was time and dose dependent. At 2 nM rhGH tyrosine phosphorylation of these two proteins was evident by 5 min, maximal at 15 min, and decreasing by 45 min of treatment. At doses of 10 and 100 nM rhGH, tyrosine phosphorylation was stimulated by 1 min of GH treatment. IM-9 cells were also treated with genetically engineered mutant forms of the GH protein. Previous biophysical analysis of these mutant GH proteins has shown that the GH protein contains two distinct binding sites which interact in a sequential manner with the extracellular domains of two distinct GH receptor molecules, thus forming a dimeric complex. By treating IM-9 cells with these same GH mutants and analyzing tyrosine phosphorylation, we found that tyrosine phosphorylation was inhibited under conditions which prevent receptor dimerization, thus providing evidence that formation of a dimeric GH:(GH receptor)2 complex may be important for intracellular signaling by the GH receptor.

This article has been cited by other articles:

				N. Yang, Y. Huang, J. Jiang, and S. J. Frank Caveolar and Lipid Raft Localization of the Growth Hormone Receptor and Its Signaling Elements: IMPACT ON GROWTH HORMONE SIGNALING J. Biol. Chem., May 14, 2004; 279(20): 20898 - 20905. [Abstract] [Full Text] [PDF]

				C. M. Silva, M. T. Kloth, A. J. Whatmore, J. S. Freeth, N. Anderson, K. K. Laughlin, T. Huynh, A. J. Woodall, and P. E. Clayton GH and Epidermal Growth Factor Signaling in Normal and Laron Syndrome Fibroblasts Endocrinology, July 1, 2002; 143(7): 2610 - 2617. [Abstract] [Full Text] [PDF]

				S. J. Frank Minireview: Receptor Dimerization in GH and Erythropoietin Action--It Takes Two to Tango, But How? Endocrinology, January 1, 2002; 143(1): 2 - 10. [Abstract] [Full Text] [PDF]

				L. Rui, S. F. Archer, L. S. Argetsinger, and C. Carter-Su Platelet-derived Growth Factor and Lysophosphatidic Acid Inhibit Growth Hormone Binding and Signaling via a Protein Kinase C-dependent Pathway J. Biol. Chem., January 28, 2000; 275(4): 2885 - 2892. [Abstract] [Full Text] [PDF]

				Y. SEGEV, D. LANDAU, R. RASCH, A. FLYVBJERG, and M. PHILLIP Growth Hormone Receptor Antagonism Prevents Early Renal in Nonobese Diabetic Mice J. Am. Soc. Nephrol., November 1, 1999; 10(11): 2374 - 2381. [Abstract] [Full Text]

				M. Maamra, J. Finidori, S. Von Laue, S. Simon, S. Justice, J. Webster, S. Dower, and R. Ross Studies with a Growth Hormone Antagonist and Dual-fluorescent Confocal Microscopy Demonstrate that the Full-length Human Growth Hormone Receptor, but Not the Truncated Isoform, Is Very Rapidly Internalized Independent of Jak2-Stat5 Signaling J. Biol. Chem., May 21, 1999; 274(21): 14791 - 14798. [Abstract] [Full Text] [PDF]

				E Juarez-Aguilar, F Castro-Munozledo, N. Guerra-Rodriguez, D Resendez-Perez, H. Martinez-Rodriguez, H. Barrera-Saldana, and W Kuri-Harcuch Functional domains of human growth hormone necessary for the adipogenic activity of hGH/hPL chimeric molecules J. Cell Sci., January 9, 1999; 112(18): 3127 - 3135. [Abstract] [PDF]

				D. W. Love, A. J. Whatmore, P. E. Clayton, and C. M. Silva Growth Hormone Stimulation of the Mitogen-Activated Protein Kinase Pathway Is Cell Type Specific Endocrinology, April 1, 1998; 139(4): 1965 - 1971. [Abstract] [Full Text] [PDF]

				S.-O. Kim, J. Jiang, W. Yi, G.-S. Feng, and S. J. Frank Involvement of the Src Homology 2-containing Tyrosine Phosphatase SHP-2 in Growth Hormone Signaling J. Biol. Chem., January 23, 1998; 273(4): 2344 - 2354. [Abstract] [Full Text] [PDF]

				C. A. Gebert, S.-H. Park, and D. J. Waxman Regulation of Signal Transducer and Activator of Transcription (STAT) 5b Activation by the Temporal Pattern of Growth Hormone Stimulation Mol. Endocrinol., April 1, 1997; 11(4): 400 - 414. [Abstract] [Full Text]

				C. Chen, R. Brinkworth, and M. J. Waters The Role of Receptor Dimerization Domain Residues in Growth Hormone Signaling J. Biol. Chem., February 21, 1997; 272(8): 5133 - 5140. [Abstract] [Full Text] [PDF]

				G. J. Strous, P. van Kerkhof, R. Govers, P. Rotwein, and A. L. Schwartz Growth Hormone-induced Signal Transduction Depends on an Intact Ubiquitin System J. Biol. Chem., January 3, 1997; 272(1): 40 - 43. [Abstract] [Full Text] [PDF]

				B. C. Xu, X. Wang, C. J. Darus, and J. J. Kopchick Growth Hormone Promotes the Association of Transcription Factor STAT5 with the Growth Hormone Receptor J. Biol. Chem., August 16, 1996; 271(33): 19768 - 19773. [Abstract] [Full Text] [PDF]

				P. A. Harding, X. Wang, S. Okada, W. Y. Chen, W. Wan, and J. J. Kopchick Growth Hormone (GH) and a GH Antagonist Promote GH Receptor Dimerization and Internalization J. Biol. Chem., March 22, 1996; 271(12): 6708 - 6712. [Abstract] [Full Text] [PDF]

				Y. Takahashi, H. Kaji, Y. Okimura, K. Goji, H. Abe, and K. Chihara Short Stature Caused by a Mutant Growth Hormone N. Engl. J. Med., February 15, 1996; 334(7): 432 - 436. [Full Text] [PDF]

				R. H. Hackett, Y.-D. Wang, and A. C. Larner Mapping of the Cytoplasmic Domain of the Human Growth Hormone Receptor Required for the Activation of Jak2 and Stat Proteins J. Biol. Chem., September 8, 1995; 270(36): 21326 - 21330. [Abstract] [Full Text] [PDF]

				M. T. Dattani, P. C. Hindmarsh, C. G. D. Brook, I. C. A. F. Robinson, J. J. Kopchick, and N. J. Marshall G120R, a Human Growth Hormone Antagonist, Shows Zinc-dependent Agonist and Antagonist Activity on Nb2 Cells J. Biol. Chem., April 21, 1995; 270(16): 9222 - 9226. [Abstract] [Full Text] [PDF]

				Y.-D. Wang, K. Wong, and W. I. Wood Intracellular Tyrosine Residues of the Human Growth Hormone Receptor Are Not Required for the Signaling of Proliferation or Jak-STAT Activation J. Biol. Chem., March 31, 1995; 270(13): 7021 - 7024. [Abstract] [Full Text] [PDF]

				X. Wang, S. C. Souza, B. Kelder, J. A. Cioffi, and J. J. Kopchick A 40-Amino Acid Segment of the Growth Hormone Receptor Cytoplasmic Domain Is Essential for GH-induced Tyrosine-phosphorylated Cytosolic Proteins J. Biol. Chem., March 17, 1995; 270(11): 6261 - 6266. [Abstract] [Full Text] [PDF]