Complementary deoxyribonucleic acid cloning, gene expression, and ligand selectivity of a novel gonadotropin-releasing hormone receptor expressed in the pituitary and midbrain of Xenopus laevis.
We have cloned the full-length complementary DNA (cDNA) for a GnRH receptor from Xenopus laevis pituitary cDNA and determined its gene structure. The cDNA encodes a 368-amino acid protein that has a 46% amino acid identity to the human GnRH receptor. The X laevis GnRH receptor has all of the amino acids identified in the mammalian GnRH receptors as sites of interaction with the GnRH ligand. However, this receptor cDNA shares the same distinguishing structural features of the GnRH receptor that have been characterized from other nonmammalian vertebrates. These include the pair of aspartate residues in the transmembrane domains II and VII compared with the aspartate/asparagine arrangement in mammalian receptors, the amino acid PEY motif in extracellular loop III (SEP in mammals), and the presence of a carboxyl-terminal tail. Previous studies have reported that mammalian GnRH was equipotent to other naturally occurring GnRH subtypes in stimulating LH release from the amphibian pituitary. However, in this study we show that the X. laevis GnRH receptor has ligand selectivity for the naturally occurring GnRHs similar to other nonmammalian GnRH receptors. The order of potency of the GnRHs in stimulating inositol phosphate production in COS-1 cells transiently transfected with the X. laevis GnRH receptor cDNA was chicken GnRH II>salmon GnRH>mammalian GnRH. Transcripts of this GnRH receptor are expressed in the pituitary and midbrain of X. laevis.
Troskie
B E
BE
Department of Medical Biochemistry, University of Cape Town, Observatory, South Africa. Brigitte@Curie.uct.ac.za
Hapgood
J P
JP
Millar
R P
RP
Illing
N
N
D000595:Amino Acid Sequence; D000818:Animals; D018076:DNA, Complementary / Q000235:genetics*; D005786:Gene Expression Regulation; D006054:Goldfish; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008636:Mesencephalon / Q000378:metabolism*; D008969:Molecular Sequence Data; D010902:Pituitary Gland / Q000378:metabolism*; D011966:Receptors, LHRH / Q000096:biosynthesis / Q000235:genetics*; D016415:Sequence Alignment; D014982:Xenopus laevis / Q000235:genetics / Q000378:metabolism*
D018076:DNA, Complementary; D008024:Ligands; D011966:Receptors, LHRH; D007987:Gonadotropin-Releasing Hormone