Gonadotropin-releasing hormone receptors with intracellular carboxyl-terminal tails undergo acute desensitization of total inositol phosphate production and exhibit accelerated internalization kinetics

被引:117
作者
Heding, A
Vrecl, M
Bogerd, J
McGregor, A
Sellar, R
Taylor, PL
Eidne, KA
机构
[1] MRC, Ctr Reprod Biol, Reprod Biol Unit, Edinburgh EH3 9EW, Midlothian, Scotland
[2] Univ Utrecht, Dept Expt Zool, NL-3584 CH Utrecht, Netherlands
关键词
D O I
10.1074/jbc.273.19.11472
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The mammalian gonadotropin-releasing hormone receptor (GnRH-R) is the only G-protein-coupled receptor (GPCR) in which the intracellular C-terminal tail is completely absent. In contrast to other GPCRs, the GnRH-R does not show rapid desensitization of total inositol (IP) production, and the rates of internalization are exceptionally slow. We investigated whether the incorporation of a cytoplasmic tail into the C terminus of the GnRH-R affects desensitization events and receptor internalization rates. A GnRH-R/TRH-R chimera was created where the intracellular tail of the rat thyrotropin-releasing hormone receptor (TRH-R) was engineered into the C terminus of the rat GnRH-R, Three different rat GnRH-R cDNA stop codon mutations tone for each reading frame) were also made. The GnRH-stimulated IP production of the wild-type rat GnRH-R expressed in either COS-7 or HEK 293 cells did not desensitize even after prolonged stimulation with GnRH., In contrast, the catfish GnRH-R (which does possess an intracellular tail) and the TRH-R rapidly (<10 min) desensitized following agonist stimulation. The GnRH-R/TRH-R chimera also desensitized following treatment with GnRH, resembling the pattern shown by the TRH-R and the catfish GnRH-R, Two of the stop codon mutants did not show desensitization of IP production, and the third mutant with the longest tail was not functional. Internalization experiments showed that the rat GnRH-R had the slowest endocytosis and recycling rates compared with the TRH-R, the catfish GnRH-R, and the chimeric GnRH/TRH-R. This study demonstrates that the addition of a functional intracellular C-terminal tail to the GnRH-R produces rapid desensitization of IP production and significantly increases internalization rates.
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页码:11472 / 11477
页数:6
相关论文
共 26 条
[1]   CHARACTERIZATION OF THE GONADOTROPIN-RELEASING-HORMONE RECEPTOR IN ALPHA-T3-1 PITUITARY GONADOTROPH CELLS [J].
ANDERSON, L ;
MILLIGAN, G ;
EIDNE, KA .
JOURNAL OF ENDOCRINOLOGY, 1993, 136 (01) :51-&
[2]   RAPID DESENSITIZATION OF GNRH-STIMULATED INTRACELLULAR SIGNALING EVENTS IN ALPHA-T3-1 AND HEK-293 CELLS EXPRESSING THE GNRH RECEPTOR [J].
ANDERSON, L ;
MCGREGOR, A ;
COOK, JV ;
CHILVERS, E ;
EIDNE, KA .
ENDOCRINOLOGY, 1995, 136 (11) :5228-5231
[3]  
Anderson L, 1996, Rev Reprod, V1, P193, DOI 10.1530/ror.0.0010193
[4]  
ANDERSON L, 1995, BIOCHEM J, V311, P3565
[5]   Mutations of the conserved DRS motif in the second intracellular loop of the gonadotropin-releasing hormone receptor affect expression, activation, and internalization [J].
Arora, KK ;
Cheng, ZY ;
Catt, KJ .
MOLECULAR ENDOCRINOLOGY, 1997, 11 (09) :1203-1212
[6]   HUMAN PLACENTAL GONADOTROPIN-RELEASING-HORMONE (GNRH) BINDING-SITES .1. CHARACTERIZATION, PROPERTIES AND LIGAND SPECIFICITY [J].
BRAMLEY, TA ;
MCPHIE, CA ;
MENZIES, GS .
PLACENTA, 1992, 13 (06) :555-581
[7]   EFFECTS OF ASN(87) AND ASP(318) MUTATIONS ON LIGAND-BINDING AND SIGNAL-TRANSDUCTION IN THE RAT GNRH RECEPTOR [J].
COOK, JV ;
FACCENDA, E ;
ANDERSON, L ;
COUPER, GG ;
EIDNE, KA ;
TAYLOR, PL .
JOURNAL OF ENDOCRINOLOGY, 1993, 139 (03) :R1-R4
[8]   ABSENCE OF RAPID DESENSITIZATION OF TBE MOUSE GONADOTROPIN-RELEASING-HORMONE RECEPTOR [J].
DAVIDSON, JS ;
WAKEFIELD, IK ;
MILLAR, RP .
BIOCHEMICAL JOURNAL, 1994, 300 :299-302
[9]   MOLECULAR-CLONING AND CHARACTERIZATION OF THE RAT PITUITARY GONADOTROPIN-RELEASING-HORMONE (GNRH) RECEPTOR [J].
EIDNE, KA ;
SELLAR, RE ;
COUPER, G ;
ANDERSON, L ;
TAYLOR, PL .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 1992, 90 (01) :R5-R9
[10]  
FALCKPEDERSEN E, 1994, MOL PHARMACOL, V45, P684