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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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

id	todo:paper_19326203_v11_n3_pe0151862_AprileGarcia
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spelling	todo:paper_19326203_v11_n3_pe0151862_AprileGarcia2023-10-03T16:34:43Z Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function 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. 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 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. Fil:Paez-Pereda, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Liberman, A.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Gerez, J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Hoijman, E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Refojo, D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_19326203_v11_n3_pe0151862_AprileGarcia
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	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
spellingShingle	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 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. Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function
topic_facet	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
description	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.
format	JOUR
author	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.
author_facet	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.
author_sort	Aprile-Garcia, F.
title	Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function
title_short	Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function
title_full	Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function
title_fullStr	Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function
title_full_unstemmed	Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function
title_sort	co-expression of wild-type p2x7r with gln460arg variant alters receptor function
url	http://hdl.handle.net/20.500.12110/paper_19326203_v11_n3_pe0151862_AprileGarcia
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Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function

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