Splice variants of the relaxin and INSL3 receptors reveal unanticipated molecular complexity.
LGR7 and LGR8 are G protein-coupled receptors that belong to the leucine-rich repeat-containing G-protein coupled receptor (LGR) family, including the thyroid-stimulating hormone (TSH), LH and FSH receptors. LGR7 and LGR8 stimulate cAMP production upon binding of the cognate ligands, relaxin and insulin-like peptide 3 (INSL3), respectively. We cloned several novel splice variants of both LGR7 and LGR8 and analysed the function of four variants. LGR7.1 is a truncated receptor, including only the N-terminal region of the receptor and two leucine rich repeats. In contrast, LGR7.2, LGR7.10 and LGR 8.1 all contain an intact seven transmembrane domain and most of the extracellular region, lacking only one or two exons in the ectodomain. Our analysis demonstrates that although LGR7.10 and LGR8.1 are expressed at the cell surface, LGR7.2 is predominantly retained within cells and LGR7.1 is partially secreted. mRNA expression analysis revealed that several variants are co-expressed in various tissues. None of these variants were able to stimulate cAMP production following relaxin or INSL3 treatment. Unexpectedly, we did not detect any direct specific relaxin or INSL3 binding on any of the splice variants. The large number of receptor splice variants identified suggests an unforeseen complexity in the physiology of this novel hormone-receptor system.
Muda
Marco
M
Serono Research Institute, One Technology Place, Rockland, MA 02370, USA. marco.muda@serono.com
He
Chaomei
C
Martini
Paolo G V
PG
Ferraro
Tania
T
Layfield
Sharon
S
Taylor
Deanne
D
Chevrier
Colette
C
Schweickhardt
Rene
R
Kelton
Christie
C
Ryan
Peter L
PL
Bathgate
Ross A D
RA
D017398:Alternative Splicing / Q000235:genetics*; D001483:Base Sequence; D006801:Humans; D007328:Insulin / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000235:genetics* / Q000378:metabolism; D018000:Receptors, Peptide; D012065:Relaxin / Q000378:metabolism
D007328:Insulin; C084797:Leydig insulin-like protein; D008565:Membrane Proteins; D020033:Protein Isoforms; D011506:Proteins; D012333:RNA, Messenger; C417543:RXFP1 protein, human; C472867:RXFP2 protein, human; D043562:Receptors, G-Protein-Coupled; D018000:Receptors, Peptide; D012065:Relaxin