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(Received for publication, January 24, 1997, and in revised form, April 7, 1997)
From INSERM Unité 344, Endocrinologie Moléculaire,
Faculté de Médecine Necker-Enfants Malades, 156 rue de
Vaugirard, 75730 Paris Cedex 15, France and the
¶ Département de Physiologie Systémique, CERMA, Base
d'Essais en Vol, 91228 Brétigny-sur-Orge Cedex, France
The growth hormone receptor (GHR) cDNA was
cloned from the liver of Rhesus macaque using polymerase chain
reaction. As deduced from the nucleotide sequence, the mature GHR is a
protein of 620 amino acids which presents 94.1% identity with the
human receptor. The monkey GHR (mkGHR) expressed in 293 cells presented
the expected specificity for a primate GHR and was able to transduce a
transcriptional effect of GH. Human GH was able to activate tyrosine
phosphorylation of both the tyrosine kinase JAK2 and the receptor in
293 cells co-transfected with mkGHR and JAK2 cDNAs. The GH binding
protein (GHBP), the soluble short form of the GHR, was also present in monkey serum. Expression of the GHR cDNA in eucaryotic cells
indicated that the GHBP can be produced by proteolytic cleavage of the
membrane receptor. Northern blot analysis of GHR gene expression in
different tissues allowed us to identify three different transcripts of 5.0 and 2.8 kilobase pairs and a smaller one of 1.7 kilobase pairs which could encode a GHBP. Rapid amplification of cDNA extremities (3
Volume 272, Number 30,
Issue of July 25, 1997
pp. 18951-18958
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
EVIDENCE FOR TWO MECHANISMS FOR THE GENERATION OF THE GH BINDING
PROTEIN
-RACE-polymerase chain reaction) was used to identify a cDNA encoding a protein in which the transmembrane and cytoplasmic domains
of the receptor are substituted by a short sequence of 9 amino acids.
This transcript was present in various tissues and could encode a GHBP
as well, suggesting for the first time that two different mechanisms
can coexist for the generation of the GHBP: proteolytic cleavage of the
membrane receptor and a specific mRNA produced by alternative
splicing.
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