Members of RTP and REEP gene families influence functional bitter taste receptor expression.
Functional characterization of chemosensory receptors is usually achieved by heterologous expression in mammalian cell lines. However, many chemoreceptor genes, including bitter taste receptors (TAS2Rs), show only marginal cell surface expression. Usually, these problems are circumvented by using chimeric receptors consisting of "export tags" and the receptor sequence itself. It seems likely that chemoreceptor cells express factors for cell surface targeting of native receptor molecules in vivo. For TAS2Rs, however, such factors are still unknown. The present study investigates the influence of RTP and REEP proteins on the functional expression of human TAS2Rs in heterologous cells. We expressed hTAS2Rs in HEK 293T cells and observed dramatic differences in responsiveness to agonist stimulation. By immunocytochemistry we show accumulation of the bitter beta-glucopyranoside receptor hTAS2R16 in the Golgi compartment. Coexpression of RTP and REEP proteins changed the responses of some hTAS2Rs upon agonist stimulation, which is likely due to efficient cell surface localization as demonstrated by cell surface biotinylation experiments. The coimmunoprecipitation of hTAS2R16 and RTP3 or RTP4 suggests that the mechanism by which these cofactors influence hTAS2R16 function might involve direct protein-protein interaction. Finally, expression analyses demonstrate RTP and REEP gene expression in human circumvallate papillae and testis, both of which are sites of TAS2R gene expression.
Behrens
Maik
M
Department of Molecular Genetics, German Institute of Human Nutrition Potsdam-Rehbruecke, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, Germany.
Bartelt
Juliane
J
Reichling
Claudia
C
Winnig
Marcel
M
Kuhn
Christina
C
Meyerhof
Wolfgang
W
D000818:Animals; D001483:Base Sequence; D002462:Cell Membrane / Q000502:physiology; D003001:Cloning, Molecular; D006801:Humans; D008297:Male; D026901:Membrane Transport Proteins / Q000235:genetics*; D005810:Multigene Family; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D013649:Taste / Q000235:genetics*; D013650:Taste Buds / Q000502:physiology; D013737:Testis / Q000502:physiology; D014059:Tongue / Q000502:physiology
D026901:Membrane Transport Proteins; C495318:REEP1 protein, human; C495316:RTP1 protein, human; D011994:Recombinant Proteins