An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse

Autores
Almedom, Ruta B; Liewald, Jana F; Hernando, Guillermina Silvana; Schultheis, Christian; Rayes, Diego Hernán; Pan, Jie; Schedletzky, Thorsten; Hutter, Harald; Bouzat, Cecilia Beatriz; Gottschalk, Alexander
Año de publicación
2009
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Nicotinic acetylcholine receptors (nAChRs) mediate fast excitatory neurotransmission in neurons and muscles. To identify nAChR accessory proteins, which may regulate their expression or function, we performed tandem affinity purification of the levamisole-sensitive nAChR from Caenorhabditis elegans, mass spectrometry of associated components, and RNAi-based screening for effects on in vivo nicotine sensitivity. Among the proteins identified was the calcineurin A subunit TAX-6, which appeared to function as a negative regulator of nAChR activity. We also identified five proteins not previously linked to nAChR function, whose inactivation conferred nicotine resistance, implicating them as positive regulators of nAChR activity. Of these, the copine NRA-1 colocalized with the levamisole receptor at neuronal and muscle plasma membranes, and, when mutated, caused reduced synaptic nAChR expres-sion. Loss of SOC-1, which acts in receptor tyrosine kinase (RTK) signaling, also reduced synaptic levamisole recep-tor levels, as did mutations in the fibroblast growth factor receptor EGL-15, and another RTK, CAM-1. Thus, tándem affinity purification is a viable approach to identify novel proteins regulating neurotransmitter receptor activity or expression in model systems like C. elegans.
Fil: Almedom, Ruta B. Johann Wolfgang Goethe-University; Alemania
Fil: Liewald, Jana F. Johann Wolfgang Goethe-University; Alemania
Fil: Hernando, Guillermina Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Schultheis, Christian. Johann Wolfgang Goethe-University; Alemania
Fil: Rayes, Diego Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Pan, Jie. Simon Fraser University; Canadá
Fil: Schedletzky, Thorsten. Johann Wolfgang Goethe-University; Alemania
Fil: Hutter, Harald. Simon Fraser University; Canadá
Fil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Gottschalk, Alexander. Johann Wolfgang Goethe-University; Alemania. Goethe Universitat Frankfurt; Alemania
Materia
channelrhodopsin-2
nAChR biogenesis
Nicalin
single-channel properties
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/42011
Acceder
id CONICETDig_33c796efa72ec2b7fe52294b1821186d
oai_identifier_str oai:ri.conicet.gov.ar:11336/42011
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapseAlmedom, Ruta BLiewald, Jana FHernando, Guillermina SilvanaSchultheis, ChristianRayes, Diego HernánPan, JieSchedletzky, ThorstenHutter, HaraldBouzat, Cecilia BeatrizGottschalk, Alexanderchannelrhodopsin-2nAChR biogenesisNicalinsingle-channel propertieshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Nicotinic acetylcholine receptors (nAChRs) mediate fast excitatory neurotransmission in neurons and muscles. To identify nAChR accessory proteins, which may regulate their expression or function, we performed tandem affinity purification of the levamisole-sensitive nAChR from Caenorhabditis elegans, mass spectrometry of associated components, and RNAi-based screening for effects on in vivo nicotine sensitivity. Among the proteins identified was the calcineurin A subunit TAX-6, which appeared to function as a negative regulator of nAChR activity. We also identified five proteins not previously linked to nAChR function, whose inactivation conferred nicotine resistance, implicating them as positive regulators of nAChR activity. Of these, the copine NRA-1 colocalized with the levamisole receptor at neuronal and muscle plasma membranes, and, when mutated, caused reduced synaptic nAChR expres-sion. Loss of SOC-1, which acts in receptor tyrosine kinase (RTK) signaling, also reduced synaptic levamisole recep-tor levels, as did mutations in the fibroblast growth factor receptor EGL-15, and another RTK, CAM-1. Thus, tándem affinity purification is a viable approach to identify novel proteins regulating neurotransmitter receptor activity or expression in model systems like C. elegans.Fil: Almedom, Ruta B. Johann Wolfgang Goethe-University; AlemaniaFil: Liewald, Jana F. Johann Wolfgang Goethe-University; AlemaniaFil: Hernando, Guillermina Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Schultheis, Christian. Johann Wolfgang Goethe-University; AlemaniaFil: Rayes, Diego Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Pan, Jie. Simon Fraser University; CanadáFil: Schedletzky, Thorsten. Johann Wolfgang Goethe-University; AlemaniaFil: Hutter, Harald. Simon Fraser University; CanadáFil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Gottschalk, Alexander. Johann Wolfgang Goethe-University; Alemania. Goethe Universitat Frankfurt; AlemaniaNature Publishing Group2009-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/42011Almedom, Ruta B; Liewald, Jana F; Hernando, Guillermina Silvana; Schultheis, Christian; Rayes, Diego Hernán; et al.; An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse; Nature Publishing Group; Embo Journal; 28; 17; 7-2009; 2636-26490261-4189CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://emboj.embopress.org/content/28/17/2636info:eu-repo/semantics/altIdentifier/doi/10.1038%2Femboj.2009.204info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:20:07Zoai:ri.conicet.gov.ar:11336/42011instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:20:07.831CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse
title An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse
spellingShingle An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse
Almedom, Ruta B
channelrhodopsin-2
nAChR biogenesis
Nicalin
single-channel properties
title_short An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse
title_full An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse
title_fullStr An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse
title_full_unstemmed An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse
title_sort An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse
dc.creator.none.fl_str_mv Almedom, Ruta B
Liewald, Jana F
Hernando, Guillermina Silvana
Schultheis, Christian
Rayes, Diego Hernán
Pan, Jie
Schedletzky, Thorsten
Hutter, Harald
Bouzat, Cecilia Beatriz
Gottschalk, Alexander
author Almedom, Ruta B
author_facet Almedom, Ruta B
Liewald, Jana F
Hernando, Guillermina Silvana
Schultheis, Christian
Rayes, Diego Hernán
Pan, Jie
Schedletzky, Thorsten
Hutter, Harald
Bouzat, Cecilia Beatriz
Gottschalk, Alexander
author_role author
author2 Liewald, Jana F
Hernando, Guillermina Silvana
Schultheis, Christian
Rayes, Diego Hernán
Pan, Jie
Schedletzky, Thorsten
Hutter, Harald
Bouzat, Cecilia Beatriz
Gottschalk, Alexander
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv channelrhodopsin-2
nAChR biogenesis
Nicalin
single-channel properties
topic channelrhodopsin-2
nAChR biogenesis
Nicalin
single-channel properties
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Nicotinic acetylcholine receptors (nAChRs) mediate fast excitatory neurotransmission in neurons and muscles. To identify nAChR accessory proteins, which may regulate their expression or function, we performed tandem affinity purification of the levamisole-sensitive nAChR from Caenorhabditis elegans, mass spectrometry of associated components, and RNAi-based screening for effects on in vivo nicotine sensitivity. Among the proteins identified was the calcineurin A subunit TAX-6, which appeared to function as a negative regulator of nAChR activity. We also identified five proteins not previously linked to nAChR function, whose inactivation conferred nicotine resistance, implicating them as positive regulators of nAChR activity. Of these, the copine NRA-1 colocalized with the levamisole receptor at neuronal and muscle plasma membranes, and, when mutated, caused reduced synaptic nAChR expres-sion. Loss of SOC-1, which acts in receptor tyrosine kinase (RTK) signaling, also reduced synaptic levamisole recep-tor levels, as did mutations in the fibroblast growth factor receptor EGL-15, and another RTK, CAM-1. Thus, tándem affinity purification is a viable approach to identify novel proteins regulating neurotransmitter receptor activity or expression in model systems like C. elegans.
Fil: Almedom, Ruta B. Johann Wolfgang Goethe-University; Alemania
Fil: Liewald, Jana F. Johann Wolfgang Goethe-University; Alemania
Fil: Hernando, Guillermina Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Schultheis, Christian. Johann Wolfgang Goethe-University; Alemania
Fil: Rayes, Diego Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Pan, Jie. Simon Fraser University; Canadá
Fil: Schedletzky, Thorsten. Johann Wolfgang Goethe-University; Alemania
Fil: Hutter, Harald. Simon Fraser University; Canadá
Fil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Gottschalk, Alexander. Johann Wolfgang Goethe-University; Alemania. Goethe Universitat Frankfurt; Alemania
description Nicotinic acetylcholine receptors (nAChRs) mediate fast excitatory neurotransmission in neurons and muscles. To identify nAChR accessory proteins, which may regulate their expression or function, we performed tandem affinity purification of the levamisole-sensitive nAChR from Caenorhabditis elegans, mass spectrometry of associated components, and RNAi-based screening for effects on in vivo nicotine sensitivity. Among the proteins identified was the calcineurin A subunit TAX-6, which appeared to function as a negative regulator of nAChR activity. We also identified five proteins not previously linked to nAChR function, whose inactivation conferred nicotine resistance, implicating them as positive regulators of nAChR activity. Of these, the copine NRA-1 colocalized with the levamisole receptor at neuronal and muscle plasma membranes, and, when mutated, caused reduced synaptic nAChR expres-sion. Loss of SOC-1, which acts in receptor tyrosine kinase (RTK) signaling, also reduced synaptic levamisole recep-tor levels, as did mutations in the fibroblast growth factor receptor EGL-15, and another RTK, CAM-1. Thus, tándem affinity purification is a viable approach to identify novel proteins regulating neurotransmitter receptor activity or expression in model systems like C. elegans.
publishDate 2009
dc.date.none.fl_str_mv 2009-07
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/42011
Almedom, Ruta B; Liewald, Jana F; Hernando, Guillermina Silvana; Schultheis, Christian; Rayes, Diego Hernán; et al.; An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse; Nature Publishing Group; Embo Journal; 28; 17; 7-2009; 2636-2649
0261-4189
CONICET Digital
CONICET
url http://hdl.handle.net/11336/42011
identifier_str_mv Almedom, Ruta B; Liewald, Jana F; Hernando, Guillermina Silvana; Schultheis, Christian; Rayes, Diego Hernán; et al.; An ER-resident membrane protein complex regulates nicotinic acetylcholine receptor subunit composition at the synapse; Nature Publishing Group; Embo Journal; 28; 17; 7-2009; 2636-2649
0261-4189
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://emboj.embopress.org/content/28/17/2636
info:eu-repo/semantics/altIdentifier/doi/10.1038%2Femboj.2009.204
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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