Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium

Autores
Olivera Couto, Agustina; Salzman, Valentina; Mailhos, Milagros; Digman, Michelle A.; Gratton, Enrico; Aguilar, Pablo Sebastián
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Eisosomes are plasma membrane domains concentrating lipids, transporters, and signaling molecules. In the budding yeast Saccharomyces cerevisiae, these domains are structured by scaffolds composed mainly by two cytoplasmic proteins Pil1 and Lsp1. Eisosomes are immobile domains, have relatively uniform size, and encompass thousands of units of the core proteins Pil1 and Lsp1. In this work we used fluorescence fluctuation analytical methods to determine the dynamics of eisosome core proteins at different subcellular locations. Using a combination of scanning techniques with autocorrelation analysis, we show that Pil1 and Lsp1 cytoplasmic pools freely diffuse whereas an eisosome-associated fraction of these proteins exhibits slow dynamics that fit with a binding-unbinding equilibrium. Number and brightness analysis shows that the eisosome-associated fraction is oligomeric, while cytoplasmic pools have lower aggregation states. Fluorescence lifetime imaging results indicate that Pil1 and Lsp1 directly interact in the cytoplasm and within the eisosomes. These results support a model where Pil1-Lsp1 heterodimers are the minimal eisosomes building blocks. Moreover, individual-eisosome fluorescence fluctuation analysis shows that eisosomes in the same cell are not equal domains: while roughly half of them are mostly static, the other half is actively exchanging core protein subunits.
Fil: Olivera Couto, Agustina. Instituto Pasteur de Montevideo; Uruguay
Fil: Salzman, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Instituto Pasteur de Montevideo; Uruguay
Fil: Mailhos, Milagros. Instituto Pasteur de Montevideo; Uruguay
Fil: Digman, Michelle A.. University of California at Irvine; Estados Unidos. University of New England; Australia
Fil: Gratton, Enrico. University of California at Irvine; Estados Unidos
Fil: Aguilar, Pablo Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Instituto Pasteur de Montevideo; Uruguay
Materia
Nanodominios
Mmembrana plasmática
Llevaduras
Microscopía
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/49343
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/49343
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibriumOlivera Couto, AgustinaSalzman, ValentinaMailhos, MilagrosDigman, Michelle A.Gratton, EnricoAguilar, Pablo SebastiánNanodominiosMmembrana plasmáticaLlevadurasMicroscopíahttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Eisosomes are plasma membrane domains concentrating lipids, transporters, and signaling molecules. In the budding yeast Saccharomyces cerevisiae, these domains are structured by scaffolds composed mainly by two cytoplasmic proteins Pil1 and Lsp1. Eisosomes are immobile domains, have relatively uniform size, and encompass thousands of units of the core proteins Pil1 and Lsp1. In this work we used fluorescence fluctuation analytical methods to determine the dynamics of eisosome core proteins at different subcellular locations. Using a combination of scanning techniques with autocorrelation analysis, we show that Pil1 and Lsp1 cytoplasmic pools freely diffuse whereas an eisosome-associated fraction of these proteins exhibits slow dynamics that fit with a binding-unbinding equilibrium. Number and brightness analysis shows that the eisosome-associated fraction is oligomeric, while cytoplasmic pools have lower aggregation states. Fluorescence lifetime imaging results indicate that Pil1 and Lsp1 directly interact in the cytoplasm and within the eisosomes. These results support a model where Pil1-Lsp1 heterodimers are the minimal eisosomes building blocks. Moreover, individual-eisosome fluorescence fluctuation analysis shows that eisosomes in the same cell are not equal domains: while roughly half of them are mostly static, the other half is actively exchanging core protein subunits.Fil: Olivera Couto, Agustina. Instituto Pasteur de Montevideo; UruguayFil: Salzman, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Instituto Pasteur de Montevideo; UruguayFil: Mailhos, Milagros. Instituto Pasteur de Montevideo; UruguayFil: Digman, Michelle A.. University of California at Irvine; Estados Unidos. University of New England; AustraliaFil: Gratton, Enrico. University of California at Irvine; Estados UnidosFil: Aguilar, Pablo Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Instituto Pasteur de Montevideo; UruguayCell Press2015-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/49343Olivera Couto, Agustina; Salzman, Valentina; Mailhos, Milagros; Digman, Michelle A.; Gratton, Enrico; et al.; Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium; Cell Press; Biophysical Journal; 108; 7; 4-2015; 1633-16440006-3495CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S000634951500171Xinfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.bpj.2015.02.011info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4390835/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:51:03Zoai:ri.conicet.gov.ar:11336/49343instacron: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 09:51:03.91CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium
title Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium
spellingShingle Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium
Olivera Couto, Agustina
Nanodominios
Mmembrana plasmática
Llevaduras
Microscopía
title_short Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium
title_full Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium
title_fullStr Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium
title_full_unstemmed Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium
title_sort Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium
dc.creator.none.fl_str_mv Olivera Couto, Agustina
Salzman, Valentina
Mailhos, Milagros
Digman, Michelle A.
Gratton, Enrico
Aguilar, Pablo Sebastián
author Olivera Couto, Agustina
author_facet Olivera Couto, Agustina
Salzman, Valentina
Mailhos, Milagros
Digman, Michelle A.
Gratton, Enrico
Aguilar, Pablo Sebastián
author_role author
author2 Salzman, Valentina
Mailhos, Milagros
Digman, Michelle A.
Gratton, Enrico
Aguilar, Pablo Sebastián
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Nanodominios
Mmembrana plasmática
Llevaduras
Microscopía
topic Nanodominios
Mmembrana plasmática
Llevaduras
Microscopía
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Eisosomes are plasma membrane domains concentrating lipids, transporters, and signaling molecules. In the budding yeast Saccharomyces cerevisiae, these domains are structured by scaffolds composed mainly by two cytoplasmic proteins Pil1 and Lsp1. Eisosomes are immobile domains, have relatively uniform size, and encompass thousands of units of the core proteins Pil1 and Lsp1. In this work we used fluorescence fluctuation analytical methods to determine the dynamics of eisosome core proteins at different subcellular locations. Using a combination of scanning techniques with autocorrelation analysis, we show that Pil1 and Lsp1 cytoplasmic pools freely diffuse whereas an eisosome-associated fraction of these proteins exhibits slow dynamics that fit with a binding-unbinding equilibrium. Number and brightness analysis shows that the eisosome-associated fraction is oligomeric, while cytoplasmic pools have lower aggregation states. Fluorescence lifetime imaging results indicate that Pil1 and Lsp1 directly interact in the cytoplasm and within the eisosomes. These results support a model where Pil1-Lsp1 heterodimers are the minimal eisosomes building blocks. Moreover, individual-eisosome fluorescence fluctuation analysis shows that eisosomes in the same cell are not equal domains: while roughly half of them are mostly static, the other half is actively exchanging core protein subunits.
Fil: Olivera Couto, Agustina. Instituto Pasteur de Montevideo; Uruguay
Fil: Salzman, Valentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Instituto Pasteur de Montevideo; Uruguay
Fil: Mailhos, Milagros. Instituto Pasteur de Montevideo; Uruguay
Fil: Digman, Michelle A.. University of California at Irvine; Estados Unidos. University of New England; Australia
Fil: Gratton, Enrico. University of California at Irvine; Estados Unidos
Fil: Aguilar, Pablo Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina. Instituto Pasteur de Montevideo; Uruguay
description Eisosomes are plasma membrane domains concentrating lipids, transporters, and signaling molecules. In the budding yeast Saccharomyces cerevisiae, these domains are structured by scaffolds composed mainly by two cytoplasmic proteins Pil1 and Lsp1. Eisosomes are immobile domains, have relatively uniform size, and encompass thousands of units of the core proteins Pil1 and Lsp1. In this work we used fluorescence fluctuation analytical methods to determine the dynamics of eisosome core proteins at different subcellular locations. Using a combination of scanning techniques with autocorrelation analysis, we show that Pil1 and Lsp1 cytoplasmic pools freely diffuse whereas an eisosome-associated fraction of these proteins exhibits slow dynamics that fit with a binding-unbinding equilibrium. Number and brightness analysis shows that the eisosome-associated fraction is oligomeric, while cytoplasmic pools have lower aggregation states. Fluorescence lifetime imaging results indicate that Pil1 and Lsp1 directly interact in the cytoplasm and within the eisosomes. These results support a model where Pil1-Lsp1 heterodimers are the minimal eisosomes building blocks. Moreover, individual-eisosome fluorescence fluctuation analysis shows that eisosomes in the same cell are not equal domains: while roughly half of them are mostly static, the other half is actively exchanging core protein subunits.
publishDate 2015
dc.date.none.fl_str_mv 2015-04
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/11336/49343
Olivera Couto, Agustina; Salzman, Valentina; Mailhos, Milagros; Digman, Michelle A.; Gratton, Enrico; et al.; Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium; Cell Press; Biophysical Journal; 108; 7; 4-2015; 1633-1644
0006-3495
CONICET Digital
CONICET
url http://hdl.handle.net/11336/49343
identifier_str_mv Olivera Couto, Agustina; Salzman, Valentina; Mailhos, Milagros; Digman, Michelle A.; Gratton, Enrico; et al.; Eisosomes are dynamic plasma membrane domains showing Pil1-Lsp1 heteroligomer binding equilibrium; Cell Press; Biophysical Journal; 108; 7; 4-2015; 1633-1644
0006-3495
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://www.sciencedirect.com/science/article/pii/S000634951500171X
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bpj.2015.02.011
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4390835/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Cell Press
publisher.none.fl_str_mv Cell Press
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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