Molecular cloning of a functional human thyrotropin-releasing hormone receptor.
A cDNA encoding the human thyrotropin-releasing hormone receptor (hTRH-R) was isolated from a human brain cDNA library. Screening of 1.2 million clones resulted in 2 candidates. The largest clone contained TRH-R homologous sequences starting in the third transmembrane domain and included a long 3' untranslated sequence. The smaller clone contained a potential start of the open reading frame, but was interrupted by an intron in the sixth transmembrane domain. The two clones had 497 bp of overlapping identical sequences and it was possible to assemble a complete cDNA thus restoring the assumed coding sequence. Electrophysiological studies of frog oocytes injected with in vitro transcribed mRNA showed TRH-specific inward currents, demonstrating that the reconstituted cDNA encoded a functional receptor. The predicted amino acid sequence of the hTRH-R protein showed high homology with the rat and mouse TRH-Rs with the exception of their C-terminal region. The human TRH-R gene seems to contain two introns.
Matre
V
V
Institute of Medical Biochemistry, University of Oslo, Norway.
Karlsen
H E
HE
Wright
M S
MS
Lundell
I
I
Fjeldheim
A K
AK
Gabrielsen
O S
OS
Larhammar
D
D
Gautvik
K M
KM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D015723:Gene Library; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D009865:Oocytes / Q000187:drug effects / Q000502:physiology; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000378:metabolism; D017981:Receptors, Neurotransmitter / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D018025:Receptors, Thyrotropin-Releasing Hormone; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D013973:Thyrotropin-Releasing Hormone / Q000378:metabolism / Q000494:pharmacology*; D014158:Transcription, Genetic; D014982:Xenopus laevis
D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D017981:Receptors, Neurotransmitter; D018025:Receptors, Thyrotropin-Releasing Hormone; D013973:Thyrotropin-Releasing Hormone; D004247:DNA