Characterization of Gpr101 expression and G-protein coupling selectivity.
This report describes the identification and characterization of the murine orphan GPCR, Gpr101. Both human and murine genes were localized to chromosome X. Similar to its human ortholog, murine Gpr101 mRNA was detected predominantly in the brain within discrete nuclei. A knowledge-restricted hidden Markov model-based algorithm, capable of accurately predicting G-protein coupling selectivity, indicated that both human and murine GPR101 were likely coupled to Gs. This prediction was supported by the elevation of cyclic AMP levels and the activation of a cyclic AMP response element-luciferase reporter gene in HEK293 cells over-expressing human GPR101. Consistent with this, over-expression of human GPR101 in a yeast-based system yielded an elevated, agonist-independent reporter gene response in the presence of a yeast chimeric Galphas protein. These results indicate that GPR101 participates in a potentially wide range of activities in the CNS via modulation of cAMP levels.
Bates
Brian
B
Wyeth Research, Biological Technologies, 87 Cambridge Park Drive, Cambridge, MA 02140, USA. bbates@wyeth.com
Zhang
Lynn
L
Nawoschik
Stan
S
Kodangattil
Sreekumar
S
Tseng
Eugene
E
Kopsco
David
D
Kramer
Angela
A
Shan
Qin
Q
Taylor
Noël
N
Johnson
Jeremy
J
Sun
Ying
Y
Chen
Hui Min
HM
Blatcher
Maria
M
Paulsen
Janet E
JE
Pausch
Mark H
MH
D000818:Animals; D015152:Blotting, Northern / Q000379:methods; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D002874:Chromosome Mapping / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods; D000242:Cyclic AMP / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000502:physiology*; D015870:Gene Expression / Q000502:physiology*; D015723:Gene Library; D017930:Genes, Reporter / Q000502:physiology; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D051379:Mice; D008954:Models, Biological; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000502:physiology*; D043562:Receptors, G-Protein-Coupled / Q000502:physiology*; D014162:Transfection / Q000379:methods; D020798:Two-Hybrid System Techniques
C518867:GPCR101 protein, mouse; D009419:Nerve Tissue Proteins; D043562:Receptors, G-Protein-Coupled; D000242:Cyclic AMP; D019204:GTP-Binding Proteins