Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function

The P2X7 receptor is a member of the P2X family of ligand-gated ion channels. A single-nucleotide polymorphism leading to a glutamine (Gln) by arginine (Arg) substitution at codon 460 of the purinergic P2X7 receptor (P2X7R) has been associated with mood disorders. No change in function (loss or gain...

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Detalles Bibliográficos
Autores principales: Aprile-Garcia, F., Metzger, M.W., Paez-Pereda, M., Stadler, H., Acuña, M., Liberman, A.C., Senin, S.A., Gerez, J., Hoijman, E., Refojo, D., Mitkovski, M., Panhuysen, M., Stühmer, W., Holsboer, F., Deussing, J.M., Arzt, E.
Formato: JOUR
Materias:
calcium
P2XR receptor, human
purinergic P2X7 receptor
small interfering RNA
fluorescence resonance energy transfer
genetics
HEK293 cell line
human
immunoprecipitation
metabolism
patch clamp technique
physiology
real time polymerase chain reaction
signal transduction
single nucleotide polymorphism
Western blotting
Blotting, Western
Calcium
Fluorescence Resonance Energy Transfer
HEK293 Cells
Humans
Immunoprecipitation
Patch-Clamp Techniques
Polymorphism, Single Nucleotide
Real-Time Polymerase Chain Reaction
Receptors, Purinergic P2X7
RNA, Small Interfering
Signal Transduction
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_19326203_v11_n3_pe0151862_AprileGarcia
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
Descripción
Sumario:The P2X7 receptor is a member of the P2X family of ligand-gated ion channels. A single-nucleotide polymorphism leading to a glutamine (Gln) by arginine (Arg) substitution at codon 460 of the purinergic P2X7 receptor (P2X7R) has been associated with mood disorders. No change in function (loss or gain) has been described for this SNP so far. Here we show that although the P2X7R-Gln460Arg variant per se is not compromised in its function, co-expression of wild-type P2X7R with P2X7R-Gln460Arg impairs receptor function with respect to calcium influx, channel currents and intracellular signaling in vitro. Moreover, co-immunoprecipitation and FRET studies show that the P2X7R-Gln460Arg variant physically interacts with P2X7R-WT. Specific silencing of either the normal or polymorphic variant rescues the heterozygous loss of function phenotype and restores normal function. The described loss of function due to co-expression, unique for mutations in the P2RX7 gene so far, explains the mechanism by which the P2X7R-Gln460Arg variant affects the normal function of the channel and may represent a mechanism of action for other mutations.

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