Evolutionary divergence of the truncated growth hormone receptor isoform in its ability to generate a soluble growth hormone binding protein.
The soluble growth hormone binding protein (GHBP), which is encoded by the GH receptor (GHR) gene, is generated by several mechanisms. In rabbits (rb) and humans (h), it is derived by proteolytic cleavage of the full-length membrane-bound receptor molecules (GHR-fl), whereas in rats (r) and mice, it results from an alternative splice excluding the transmembrane domain. Furthermore, in all these species, alternative splicing in the cytoplasmic domain results in a truncated isoform (GHR-tr), that, in humans, produces large amounts of GHBP through proteolysis. To further characterize the species specificity of the mechanism underlying GHBP generation, rbGHR-tr and rGHR-tr expressed in COS-7 cells were assayed for their ability to produce a GHBP in comparison with the corresponding full-length receptors. Large amounts of GHBP were secreted by cells expressing the rabbit constructs, the rbGHR-tr isoform being more efficient in GHBP generation than rbGHR-fl. In contrast, no GHBP was detected from cells expressing rGHR-tr, the cytoplasmic deletion having no effect on GHBP release from membrane receptors. These data further demonstrate evolutionary divergence in the mechanism by which GHBP is generated and provide new clues to decipher the molecular process underlying the cleavage step.
Dastot
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F
Laboratoire de Génétique moléculaire, Institut National de la Santé et de la Recherche Médicale U, 468, Hôpital Henri Mondor, Créteil, France.
Duquesnoy
P
P
Sobrier
M L
ML
Goossens
M
M
Amselem
S
S
D000273:Adipose Tissue / Q000378:metabolism; D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D019143:Evolution, Molecular; D005822:Genetic Vectors / Q000378:metabolism; D013006:Growth Hormone / Q000378:metabolism*; D011485:Protein Binding; D020033:Protein Isoforms / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D011817:Rabbits; D051381:Rats; D011986:Receptors, Somatotropin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D012995:Solubility
D020033:Protein Isoforms; D011986:Receptors, Somatotropin; D011994:Recombinant Proteins; D013006:Growth Hormone