Cloning and characterization of additional members of the G protein-coupled receptor family.
A search of the expressed sequence tag (EST) database retrieved a human cDNA sequence which partially encoded a novel G protein-coupled receptor (GPCR) GPR26. A human genomic DNA fragment encoding a partial open reading frame (ORF) and a rat cDNA encoding the full length ORF of GPR26 were obtained by library screening. The rat GPR26 cDNA encoded a protein of 317 amino acids, most similar (albeit distantly related) to the serotonin 5-HT(5A) and gastrin releasing hormone BB2 receptors. GPR26 mRNA expression analysis revealed signals in the striatum, pons, cerebellum and cortex. HEK293 and Rh7777 cells transfected with GPR26 cDNA displayed high basal cAMP levels, slow growth rate of clonal populations and derangements of normal cell shape. We also used a sequence reported only in the patent literature encoding GPR57 (a.k.a. HNHCI32) to PCR amplify a DNA fragment which was used to screen a human genomic library. This resulted in the cloning of a genomic fragment containing a pseudogene, psiGPR57, with a 99.6% nucleotide identity to GPR57. Based on shared sequence identities, the receptor encoded by GPR57 was predicted to belong to a novel subfamily of GPCRs together with GPR58 (a.k.a. phBL5, reported only in the patent literature), putative neurotransmitter receptor (PNR) and a 5-HT(4) pseudogene. Analysis of this subfamily revealed greatest identities (approximately 56%) between the receptors encoded by GPR57 and GPR58, each with shared identities of approximately 40% with PNR. Furthermore, psiGPR57, GPR58, PNR and the 5-HT(4) pseudogene were mapped in a cluster localized to chromosome 6q22-24. PNR and GPR58 were expressed in COS cells, however no specific binding was observed for various serotonin receptor-specific ligands.
Lee
D K
DK
Department of Pharmacology, University of Toronto, Medical Sciences Building, Toronto, Ont., Canada.
Lynch
K R
KR
Nguyen
T
T
Im
D S
DS
Cheng
R
R
Saldivia
V R
VR
Liu
Y
Y
Liu
I S
IS
Heng
H H
HH
Seeman
P
P
George
S R
SR
O'Dowd
B F
BF
Marchese
A
A
D015056:1-Methyl-3-isobutylxanthine; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D019556:COS Cells; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008024:Ligands; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011544:Pseudogenes; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled; D014162:Transfection; D000095383:Trace Amine-Associated Receptors
D015056:1-Methyl-3-isobutylxanthine; D018076:DNA, Complementary; D008024:Ligands; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C408787:Gpr26 protein, rat; C408786:TAAR2 protein, human; D000095383:Trace Amine-Associated Receptors; C408785:TAAR3P protein, human