Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis

Autores
Castillo Bennett, Jimena Victoria; Roggero Savoini, Carlos Marcelo; Mancifesta, Franco E.; Mayorga, Luis Segundo
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Regulated secretion is a fundamental process underlying thefunction of many cell types. In particular, acrosomal exocytosisin mammalian sperm is essential for egg fertilization. In general,exocytosis is initiated by a cytosolic calcium increase. In thisreport we show that calcium affects several factors duringhuman sperm acrosomal exocytosis. By using an antibody thatspecifically recognizes synaptotagmin VI phosphorylated at thepolybasic region of the C2B domain, we showed that a calciumdependentdephosphorylation of this protein occurred at earlystages of the acrosomal exocytosis in streptolysin O-permeabilizedsperm. We identified the phosphatase as calcineurin andshowed that the activity of this enzyme is absolutely requiredduring the early steps of the secretory process. When added tosperm, an inhibitor-insensitive, catalytically active domain ofcalcineurin was able to rescue the effect of the specific calcineurininhibitor cyclosporin A. This same domain dephosphorylatedrecombinant synaptotagmin VI C2B domain, validatingthis protein as a new substrate for calcineurin. Whensperm were treated with catalytically active calcineurin beforestimulation, exocytosis was inhibited, an effect that was rescuedby the phosphomimetic synaptotagmin VI C2B-T418E,T419Emutant domain. These observations indicate that synaptotagminmust be dephosphorylated at a specific window of time andsuggest that phosphorylated synaptotagmin has an active role atearly stages of the acrosomal exocytosis.
Fil: Castillo Bennett, Jimena Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Roggero Savoini, Carlos Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Mancifesta, Franco E.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Mayorga, Luis Segundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Materia
CALCINEURIN
SYNAPTOTAGMIN-VI
DEPHOSPHORYLATION
EXOCYTOSIS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/242917
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/242917
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal ExocytosisCastillo Bennett, Jimena VictoriaRoggero Savoini, Carlos MarceloMancifesta, Franco E.Mayorga, Luis SegundoCALCINEURINSYNAPTOTAGMIN-VIDEPHOSPHORYLATIONEXOCYTOSIShttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Regulated secretion is a fundamental process underlying thefunction of many cell types. In particular, acrosomal exocytosisin mammalian sperm is essential for egg fertilization. In general,exocytosis is initiated by a cytosolic calcium increase. In thisreport we show that calcium affects several factors duringhuman sperm acrosomal exocytosis. By using an antibody thatspecifically recognizes synaptotagmin VI phosphorylated at thepolybasic region of the C2B domain, we showed that a calciumdependentdephosphorylation of this protein occurred at earlystages of the acrosomal exocytosis in streptolysin O-permeabilizedsperm. We identified the phosphatase as calcineurin andshowed that the activity of this enzyme is absolutely requiredduring the early steps of the secretory process. When added tosperm, an inhibitor-insensitive, catalytically active domain ofcalcineurin was able to rescue the effect of the specific calcineurininhibitor cyclosporin A. This same domain dephosphorylatedrecombinant synaptotagmin VI C2B domain, validatingthis protein as a new substrate for calcineurin. Whensperm were treated with catalytically active calcineurin beforestimulation, exocytosis was inhibited, an effect that was rescuedby the phosphomimetic synaptotagmin VI C2B-T418E,T419Emutant domain. These observations indicate that synaptotagminmust be dephosphorylated at a specific window of time andsuggest that phosphorylated synaptotagmin has an active role atearly stages of the acrosomal exocytosis.Fil: Castillo Bennett, Jimena Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Roggero Savoini, Carlos Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Mancifesta, Franco E.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Mayorga, Luis Segundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaAmerican Society for Biochemistry and Molecular Biology2010-05info: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/11336/242917Castillo Bennett, Jimena Victoria; Roggero Savoini, Carlos Marcelo; Mancifesta, Franco E.; Mayorga, Luis Segundo; Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 285; 34; 5-2010; 26269-262780021-9258CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0021925820595914info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M109.095752info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:54:06Zoai:ri.conicet.gov.ar:11336/242917instacron: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 09:54:06.58CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis
title Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis
spellingShingle Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis
Castillo Bennett, Jimena Victoria
CALCINEURIN
SYNAPTOTAGMIN-VI
DEPHOSPHORYLATION
EXOCYTOSIS
title_short Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis
title_full Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis
title_fullStr Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis
title_full_unstemmed Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis
title_sort Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis
dc.creator.none.fl_str_mv Castillo Bennett, Jimena Victoria
Roggero Savoini, Carlos Marcelo
Mancifesta, Franco E.
Mayorga, Luis Segundo
author Castillo Bennett, Jimena Victoria
author_facet Castillo Bennett, Jimena Victoria
Roggero Savoini, Carlos Marcelo
Mancifesta, Franco E.
Mayorga, Luis Segundo
author_role author
author2 Roggero Savoini, Carlos Marcelo
Mancifesta, Franco E.
Mayorga, Luis Segundo
author2_role author
author
author
dc.subject.none.fl_str_mv CALCINEURIN
SYNAPTOTAGMIN-VI
DEPHOSPHORYLATION
EXOCYTOSIS
topic CALCINEURIN
SYNAPTOTAGMIN-VI
DEPHOSPHORYLATION
EXOCYTOSIS
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Regulated secretion is a fundamental process underlying thefunction of many cell types. In particular, acrosomal exocytosisin mammalian sperm is essential for egg fertilization. In general,exocytosis is initiated by a cytosolic calcium increase. In thisreport we show that calcium affects several factors duringhuman sperm acrosomal exocytosis. By using an antibody thatspecifically recognizes synaptotagmin VI phosphorylated at thepolybasic region of the C2B domain, we showed that a calciumdependentdephosphorylation of this protein occurred at earlystages of the acrosomal exocytosis in streptolysin O-permeabilizedsperm. We identified the phosphatase as calcineurin andshowed that the activity of this enzyme is absolutely requiredduring the early steps of the secretory process. When added tosperm, an inhibitor-insensitive, catalytically active domain ofcalcineurin was able to rescue the effect of the specific calcineurininhibitor cyclosporin A. This same domain dephosphorylatedrecombinant synaptotagmin VI C2B domain, validatingthis protein as a new substrate for calcineurin. Whensperm were treated with catalytically active calcineurin beforestimulation, exocytosis was inhibited, an effect that was rescuedby the phosphomimetic synaptotagmin VI C2B-T418E,T419Emutant domain. These observations indicate that synaptotagminmust be dephosphorylated at a specific window of time andsuggest that phosphorylated synaptotagmin has an active role atearly stages of the acrosomal exocytosis.
Fil: Castillo Bennett, Jimena Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Roggero Savoini, Carlos Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Mancifesta, Franco E.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Mayorga, Luis Segundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
description Regulated secretion is a fundamental process underlying thefunction of many cell types. In particular, acrosomal exocytosisin mammalian sperm is essential for egg fertilization. In general,exocytosis is initiated by a cytosolic calcium increase. In thisreport we show that calcium affects several factors duringhuman sperm acrosomal exocytosis. By using an antibody thatspecifically recognizes synaptotagmin VI phosphorylated at thepolybasic region of the C2B domain, we showed that a calciumdependentdephosphorylation of this protein occurred at earlystages of the acrosomal exocytosis in streptolysin O-permeabilizedsperm. We identified the phosphatase as calcineurin andshowed that the activity of this enzyme is absolutely requiredduring the early steps of the secretory process. When added tosperm, an inhibitor-insensitive, catalytically active domain ofcalcineurin was able to rescue the effect of the specific calcineurininhibitor cyclosporin A. This same domain dephosphorylatedrecombinant synaptotagmin VI C2B domain, validatingthis protein as a new substrate for calcineurin. Whensperm were treated with catalytically active calcineurin beforestimulation, exocytosis was inhibited, an effect that was rescuedby the phosphomimetic synaptotagmin VI C2B-T418E,T419Emutant domain. These observations indicate that synaptotagminmust be dephosphorylated at a specific window of time andsuggest that phosphorylated synaptotagmin has an active role atearly stages of the acrosomal exocytosis.
publishDate 2010
dc.date.none.fl_str_mv 2010-05
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/11336/242917
Castillo Bennett, Jimena Victoria; Roggero Savoini, Carlos Marcelo; Mancifesta, Franco E.; Mayorga, Luis Segundo; Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 285; 34; 5-2010; 26269-26278
0021-9258
CONICET Digital
CONICET
url http://hdl.handle.net/11336/242917
identifier_str_mv Castillo Bennett, Jimena Victoria; Roggero Savoini, Carlos Marcelo; Mancifesta, Franco E.; Mayorga, Luis Segundo; Calcineurin-mediated Dephosphorylation of Synaptotagmin VI Is Necessary for Acrosomal Exocytosis; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 285; 34; 5-2010; 26269-26278
0021-9258
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0021925820595914
info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M109.095752
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
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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