Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis

Autores
Bello, Oscar Daniel; Jouannot, Ouardane; Chaudhuri, Arunima; Stroeva, Ekaterina; Coleman, Jeff; Volynski, Kirill E.; Rothman, James E.; Krishnakumar, Shyam S.
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Regulated exocytosis, which underlies many intercellular signaling events, is a tightly controlled process often triggered by calcium ion(s) (Ca2+). Despite considerable insight into the central components involved, namely, the core fusion machinery [soluble N-ethylmaleimide?sensitive factor attachment protein receptor (SNARE)] and the principal Ca2+ sensor [C2-domain proteins like synaptotagmin (Syt)], the molecular mechanism of Ca2+-dependent release has been unclear. Here, we report that the Ca2+-sensitive oligomers of Syt1, a conserved structural feature among several C2-domain proteins, play a critical role in orchestrating Ca2+-coupled vesicular release. This follows from pHluorin-based imaging of single-vesicle exocytosis in pheochromocytoma (PC12) cells showing that selective disruption of Syt1 oligomerization using a structure-directed mutation (F349A) dramatically increases the normally low levels of constitutive exocytosis to effectively occlude Ca2+-stimulated release. We propose a parsimonious model whereby Ca2+-sensitive oligomers of Syt (or a similar C2-domain protein) assembled at the site of docking physically block spontaneous fusion until disrupted by Ca2+. Our data further suggest Ca2+-coupled vesicular release is triggered by removal of the inhibition, rather than by direct activation of the fusion machinery.
Fil: Bello, Oscar Daniel. University of Yale. School of Medicine; Estados Unidos. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Mendoza. Instituto de Histologia y Embriologia de Mendoza Dr. Mario H. Burgos. Grupo Vinculado de Investigacion y Desarrollo Biotecnologico Aplicado Al Diagnostico Al Ihem | Universidad Nacional de Cuyo. Facultad de Ciencias Medicas. Instituto de Histologia y Embriologia de Mendoza Dr. Mario H. Burgos. Grupo Vinculado de Investigacion y Desarrollo Biotecnologico Aplicado Al Diagnostico Al Ihem.; Argentina. University College London; Estados Unidos
Fil: Jouannot, Ouardane. University of Yale. School of Medicine; Estados Unidos
Fil: Chaudhuri, Arunima. University of Yale. School of Medicine; Estados Unidos
Fil: Stroeva, Ekaterina. University of Yale. School of Medicine; Estados Unidos
Fil: Coleman, Jeff. University of Yale. School of Medicine; Estados Unidos
Fil: Volynski, Kirill E.. University College London; Reino Unido
Fil: Rothman, James E.. University of Yale. School of Medicine; Estados Unidos. University College London; Estados Unidos
Fil: Krishnakumar, Shyam S.. University of Yale. School of Medicine; Estados Unidos. University College London; Estados Unidos
Materia
CALCIUM
PC12 CELLS
REGULATED EXOCYTOSIS
SNARE PROTEIN
SYNAPTOTAGMIN
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:11336/160123
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/160123
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Synaptotagmin oligomerization is essential for calcium control of regulated exocytosisBello, Oscar DanielJouannot, OuardaneChaudhuri, ArunimaStroeva, EkaterinaColeman, JeffVolynski, Kirill E.Rothman, James E.Krishnakumar, Shyam S.CALCIUMPC12 CELLSREGULATED EXOCYTOSISSNARE PROTEINSYNAPTOTAGMINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Regulated exocytosis, which underlies many intercellular signaling events, is a tightly controlled process often triggered by calcium ion(s) (Ca2+). Despite considerable insight into the central components involved, namely, the core fusion machinery [soluble N-ethylmaleimide?sensitive factor attachment protein receptor (SNARE)] and the principal Ca2+ sensor [C2-domain proteins like synaptotagmin (Syt)], the molecular mechanism of Ca2+-dependent release has been unclear. Here, we report that the Ca2+-sensitive oligomers of Syt1, a conserved structural feature among several C2-domain proteins, play a critical role in orchestrating Ca2+-coupled vesicular release. This follows from pHluorin-based imaging of single-vesicle exocytosis in pheochromocytoma (PC12) cells showing that selective disruption of Syt1 oligomerization using a structure-directed mutation (F349A) dramatically increases the normally low levels of constitutive exocytosis to effectively occlude Ca2+-stimulated release. We propose a parsimonious model whereby Ca2+-sensitive oligomers of Syt (or a similar C2-domain protein) assembled at the site of docking physically block spontaneous fusion until disrupted by Ca2+. Our data further suggest Ca2+-coupled vesicular release is triggered by removal of the inhibition, rather than by direct activation of the fusion machinery.Fil: Bello, Oscar Daniel. University of Yale. School of Medicine; Estados Unidos. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Mendoza. Instituto de Histologia y Embriologia de Mendoza Dr. Mario H. Burgos. Grupo Vinculado de Investigacion y Desarrollo Biotecnologico Aplicado Al Diagnostico Al Ihem | Universidad Nacional de Cuyo. Facultad de Ciencias Medicas. Instituto de Histologia y Embriologia de Mendoza Dr. Mario H. Burgos. Grupo Vinculado de Investigacion y Desarrollo Biotecnologico Aplicado Al Diagnostico Al Ihem.; Argentina. University College London; Estados UnidosFil: Jouannot, Ouardane. University of Yale. School of Medicine; Estados UnidosFil: Chaudhuri, Arunima. University of Yale. School of Medicine; Estados UnidosFil: Stroeva, Ekaterina. University of Yale. School of Medicine; Estados UnidosFil: Coleman, Jeff. University of Yale. School of Medicine; Estados UnidosFil: Volynski, Kirill E.. University College London; Reino UnidoFil: Rothman, James E.. University of Yale. School of Medicine; Estados Unidos. University College London; Estados UnidosFil: Krishnakumar, Shyam S.. University of Yale. School of Medicine; Estados Unidos. University College London; Estados UnidosNational Academy of Sciences2018-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/160123Bello, Oscar Daniel; Jouannot, Ouardane; Chaudhuri, Arunima; Stroeva, Ekaterina; Coleman, Jeff; et al.; Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis; National Academy of Sciences; Proceedings of the National Academy of Sciences of The United States of America; 115; 32; 8-2018; 7624-76310027-8424CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1073/pnas.1808792115info:eu-repo/semantics/altIdentifier/url/https://www.pnas.org/doi/pdf/10.1073/pnas.1808792115info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:59:48Zoai:ri.conicet.gov.ar:11336/160123instacron: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:34982025-09-03 09:59:48.679CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis
title Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis
spellingShingle Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis
Bello, Oscar Daniel
CALCIUM
PC12 CELLS
REGULATED EXOCYTOSIS
SNARE PROTEIN
SYNAPTOTAGMIN
title_short Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis
title_full Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis
title_fullStr Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis
title_full_unstemmed Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis
title_sort Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis
dc.creator.none.fl_str_mv Bello, Oscar Daniel
Jouannot, Ouardane
Chaudhuri, Arunima
Stroeva, Ekaterina
Coleman, Jeff
Volynski, Kirill E.
Rothman, James E.
Krishnakumar, Shyam S.
author Bello, Oscar Daniel
author_facet Bello, Oscar Daniel
Jouannot, Ouardane
Chaudhuri, Arunima
Stroeva, Ekaterina
Coleman, Jeff
Volynski, Kirill E.
Rothman, James E.
Krishnakumar, Shyam S.
author_role author
author2 Jouannot, Ouardane
Chaudhuri, Arunima
Stroeva, Ekaterina
Coleman, Jeff
Volynski, Kirill E.
Rothman, James E.
Krishnakumar, Shyam S.
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CALCIUM
PC12 CELLS
REGULATED EXOCYTOSIS
SNARE PROTEIN
SYNAPTOTAGMIN
topic CALCIUM
PC12 CELLS
REGULATED EXOCYTOSIS
SNARE PROTEIN
SYNAPTOTAGMIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Regulated exocytosis, which underlies many intercellular signaling events, is a tightly controlled process often triggered by calcium ion(s) (Ca2+). Despite considerable insight into the central components involved, namely, the core fusion machinery [soluble N-ethylmaleimide?sensitive factor attachment protein receptor (SNARE)] and the principal Ca2+ sensor [C2-domain proteins like synaptotagmin (Syt)], the molecular mechanism of Ca2+-dependent release has been unclear. Here, we report that the Ca2+-sensitive oligomers of Syt1, a conserved structural feature among several C2-domain proteins, play a critical role in orchestrating Ca2+-coupled vesicular release. This follows from pHluorin-based imaging of single-vesicle exocytosis in pheochromocytoma (PC12) cells showing that selective disruption of Syt1 oligomerization using a structure-directed mutation (F349A) dramatically increases the normally low levels of constitutive exocytosis to effectively occlude Ca2+-stimulated release. We propose a parsimonious model whereby Ca2+-sensitive oligomers of Syt (or a similar C2-domain protein) assembled at the site of docking physically block spontaneous fusion until disrupted by Ca2+. Our data further suggest Ca2+-coupled vesicular release is triggered by removal of the inhibition, rather than by direct activation of the fusion machinery.
Fil: Bello, Oscar Daniel. University of Yale. School of Medicine; Estados Unidos. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Mendoza. Instituto de Histologia y Embriologia de Mendoza Dr. Mario H. Burgos. Grupo Vinculado de Investigacion y Desarrollo Biotecnologico Aplicado Al Diagnostico Al Ihem | Universidad Nacional de Cuyo. Facultad de Ciencias Medicas. Instituto de Histologia y Embriologia de Mendoza Dr. Mario H. Burgos. Grupo Vinculado de Investigacion y Desarrollo Biotecnologico Aplicado Al Diagnostico Al Ihem.; Argentina. University College London; Estados Unidos
Fil: Jouannot, Ouardane. University of Yale. School of Medicine; Estados Unidos
Fil: Chaudhuri, Arunima. University of Yale. School of Medicine; Estados Unidos
Fil: Stroeva, Ekaterina. University of Yale. School of Medicine; Estados Unidos
Fil: Coleman, Jeff. University of Yale. School of Medicine; Estados Unidos
Fil: Volynski, Kirill E.. University College London; Reino Unido
Fil: Rothman, James E.. University of Yale. School of Medicine; Estados Unidos. University College London; Estados Unidos
Fil: Krishnakumar, Shyam S.. University of Yale. School of Medicine; Estados Unidos. University College London; Estados Unidos
description Regulated exocytosis, which underlies many intercellular signaling events, is a tightly controlled process often triggered by calcium ion(s) (Ca2+). Despite considerable insight into the central components involved, namely, the core fusion machinery [soluble N-ethylmaleimide?sensitive factor attachment protein receptor (SNARE)] and the principal Ca2+ sensor [C2-domain proteins like synaptotagmin (Syt)], the molecular mechanism of Ca2+-dependent release has been unclear. Here, we report that the Ca2+-sensitive oligomers of Syt1, a conserved structural feature among several C2-domain proteins, play a critical role in orchestrating Ca2+-coupled vesicular release. This follows from pHluorin-based imaging of single-vesicle exocytosis in pheochromocytoma (PC12) cells showing that selective disruption of Syt1 oligomerization using a structure-directed mutation (F349A) dramatically increases the normally low levels of constitutive exocytosis to effectively occlude Ca2+-stimulated release. We propose a parsimonious model whereby Ca2+-sensitive oligomers of Syt (or a similar C2-domain protein) assembled at the site of docking physically block spontaneous fusion until disrupted by Ca2+. Our data further suggest Ca2+-coupled vesicular release is triggered by removal of the inhibition, rather than by direct activation of the fusion machinery.
publishDate 2018
dc.date.none.fl_str_mv 2018-08
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/11336/160123
Bello, Oscar Daniel; Jouannot, Ouardane; Chaudhuri, Arunima; Stroeva, Ekaterina; Coleman, Jeff; et al.; Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis; National Academy of Sciences; Proceedings of the National Academy of Sciences of The United States of America; 115; 32; 8-2018; 7624-7631
0027-8424
CONICET Digital
CONICET
url http://hdl.handle.net/11336/160123
identifier_str_mv Bello, Oscar Daniel; Jouannot, Ouardane; Chaudhuri, Arunima; Stroeva, Ekaterina; Coleman, Jeff; et al.; Synaptotagmin oligomerization is essential for calcium control of regulated exocytosis; National Academy of Sciences; Proceedings of the National Academy of Sciences of The United States of America; 115; 32; 8-2018; 7624-7631
0027-8424
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1073/pnas.1808792115
info:eu-repo/semantics/altIdentifier/url/https://www.pnas.org/doi/pdf/10.1073/pnas.1808792115
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
dc.publisher.none.fl_str_mv National Academy of Sciences
publisher.none.fl_str_mv National Academy of Sciences
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 13.13397

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