Identification of Phenylalanine 346 in the Rat Growth Hormone Receptor as Being Critical for Ligand-mediated Internalization and Down-regulation (∗)

The functional significance of growth hormone (GH) receptor (GHR) internalization is unknown; therefore, we have analyzed domains and individual amino acids in the cytoplasmic region of the rat GHR required for ligand-mediated receptor internalization, receptor down-regulation, and transcriptional s...

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Bibliographic Details
Published in:The Journal of biological chemistry 1995-07, Vol.270 (29), p.17210-17214
Main Authors: Allevato, Giovanna, Billestrup, Nils, Goujon, Laure, Galsgaard, Elisabeth D., Norstedt, Gunnar, Postel-Vinay, Marie-Catherine, Kelly, Paul A., Nielsen, Jens H.
Format: Article
Language:English
Subjects:
Animals
Cells, Cultured
Down-Regulation
Growth Hormone - metabolism
Ligands
Medicin och hälsovetenskap
Phenylalanine
Rats
Receptors, Somatotropin - analysis
Receptors, Somatotropin - chemistry
Receptors, Somatotropin - metabolism
Signal Transduction
Transcription, Genetic
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Summary:The functional significance of growth hormone (GH) receptor (GHR) internalization is unknown; therefore, we have analyzed domains and individual amino acids in the cytoplasmic region of the rat GHR required for ligand-mediated receptor internalization, receptor down-regulation, and transcriptional signaling. When various mutated GHR cDNAs were transfected stably into Chinese hamster ovary cells or transiently into monkey kidney (COS-7) cells, internalization of the GHR was found to be dependent upon a domain located between amino acids 318 and 380. Mutational analysis of aromatic residues in this domain revealed that phenylalanine 346 is required for internalization. Receptor down-regulation in transiently transfected COS-7 cells was also dependent upon the phenylalanine 346 residue of the GHR, since no GH-induced down-regulation was observed in cells expressing the F346A GHR mutant. In contrast, the ability to stimulate transcription of the serine protease inhibitor 2.1 promoter by the GHR was not affected by the phenylalanine 346 to alanine mutation. These results demonstrate that phenylalanine 346 is essential for GHR internalization and down-regulation but not for transcriptional signaling, suggesting that ligand-mediated endocytosis is not a prerequisite for GH-induced gene transcription.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.270.29.17210