The signal peptide of the rat corticotropin-releasing factor receptor 1 promotes receptor expression but is not essential for establishing a functional receptor.
Approximately 5-10% of the GPCRs (G-protein-coupled receptors) contain N-terminal signal peptides that are cleaved off during receptor insertion into the ER (endoplasmic reticulum) membrane by the signal peptidases of the ER. The reason as to why only a subset of GPCRs requires these additional signal peptides is not known. We have recently shown that the signal peptide of the human ET(B)-R (endothelin B receptor) does not influence receptor expression but is necessary for the translocation of the receptor's N-tail across the ER membrane and thus for the establishment of a functional receptor [Köchl, Alken, Rutz, Krause, Oksche, Rosenthal and Schülein (2002) J. Biol. Chem. 277, 16131-16138]. In the present study, we show that the signal peptide of the rat CRF-R1 (corticotropin-releasing factor receptor 1) has a different function: a mutant of the CRF-R1 lacking the signal peptide was functional and displayed wild-type properties with respect to ligand binding and activation of adenylate cyclase. However, immunoblot analysis and confocal laser scanning microscopy revealed that the mutant receptor was expressed at 10-fold lower levels than the wild-type receptor. Northern-blot and in vitro transcription translation analyses precluded the possibility that the reduced receptor expression is due to decreased transcription or translation levels. Thus the signal peptide of the CRF-R1 promotes an early step of receptor biogenesis, such as targeting of the nascent chain to the ER membrane and/or the gating of the protein-conducting translocon of the ER membrane.
Alken
Martina
M
Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany.
Rutz
Claudia
C
Köchl
Robert
R
Donalies
Ute
U
Oueslati
Morad
M
Furkert
Jens
J
Wietfeld
Doreen
D
Hermosilla
Ricardo
R
Scholz
Anne
A
Beyermann
Michael
M
Rosenthal
Walter
W
Schülein
Ralf
R
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D021382:Protein Sorting Signals / Q000502:physiology*; D021381:Protein Transport; D051381:Rats; D018019:Receptors, Corticotropin-Releasing Hormone / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015854:Up-Regulation; D000099052:CRF Receptor, Type 1
D021382:Protein Sorting Signals; D018019:Receptors, Corticotropin-Releasing Hormone; D000099052:CRF Receptor, Type 1