Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome

Autores
Galigniana, Mario Daniel; Harrell, Jennifer M.; Housley, Paul R.; Patterson, Cam; Fisher, Stephen K.; Pratt, William B.
Año de publicación
2004
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Here, we have used a chimera of green fluorescent protein (GFP) and the glucocorticoid receptor (GR) to study retrograde movement of a model soluble (i.e., non-vesicle-associated) protein in axons and dendrites of cultured NT2-N neurons. It is known that in non-neuronal cells, the GFP-GR moves from cytoplasm to the nucleus in a steroid-dependent manner by a rapid, hsp90-dependent mechanism. When rapid movement is inhibited by geldanamycin (GA), a specific inhibitor of the protein chaperone hsp90, the GFP-GR translocates slowly to the nucleus by diffusion. Here we show that GFP-GR expressed in hormone-free neurons is localized in both cytoplasm and neurites, and upon treatment with dexamethasone (DEX), it moves to the nucleus. In neurites, movement by diffusion is not possible, and we show that movement of the GFP-GR from neurites is blocked by geldanamycin, suggesting that the hsp90-dependent movement machinery is required for retrograde movement. In cells treated with both dexamethasone and geldanamycin, the GFP-GR becomes concentrated in fluorescent globules located periodically along the neurites. Carboxyl terminus of Hsc70-interacting protein (CHIP), the E3 ubiquitin ligase for the GR, also concentrates in the same loci in a steroid-dependent and geldanamycin-dependent manner. If geldanamycin is removed, the GFP-GR exits the globules and continues its retrograde movement. However, in the continued presence of geldanamycin, the GFP-GR in the globules undergoes proteasomal degradation, suggesting that the globules function as degradasomes. This is the first evidence for a linkage between receptor trafficking along neurites and receptor degradation by the proteasome.
Fil: Galigniana, Mario Daniel. University of Michigan; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Harrell, Jennifer M.. University of Michigan; Estados Unidos
Fil: Housley, Paul R.. University of South Carolina; Estados Unidos
Fil: Patterson, Cam. University of North Carolina; Estados Unidos
Fil: Fisher, Stephen K.. University of Michigan; Estados Unidos
Fil: Pratt, William B.. University of Michigan; Estados Unidos
Materia
Protein Trafficking
Hsp90
Proteasomal Degradation
Glucocorticoid Receptor
Chip
Geldanamycin
Neuronal Cells
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:11336/29101

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network_name_str CONICET Digital (CONICET)
spelling Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosomeGaligniana, Mario DanielHarrell, Jennifer M.Housley, Paul R.Patterson, CamFisher, Stephen K.Pratt, William B.Protein TraffickingHsp90Proteasomal DegradationGlucocorticoid ReceptorChipGeldanamycinNeuronal Cellshttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Here, we have used a chimera of green fluorescent protein (GFP) and the glucocorticoid receptor (GR) to study retrograde movement of a model soluble (i.e., non-vesicle-associated) protein in axons and dendrites of cultured NT2-N neurons. It is known that in non-neuronal cells, the GFP-GR moves from cytoplasm to the nucleus in a steroid-dependent manner by a rapid, hsp90-dependent mechanism. When rapid movement is inhibited by geldanamycin (GA), a specific inhibitor of the protein chaperone hsp90, the GFP-GR translocates slowly to the nucleus by diffusion. Here we show that GFP-GR expressed in hormone-free neurons is localized in both cytoplasm and neurites, and upon treatment with dexamethasone (DEX), it moves to the nucleus. In neurites, movement by diffusion is not possible, and we show that movement of the GFP-GR from neurites is blocked by geldanamycin, suggesting that the hsp90-dependent movement machinery is required for retrograde movement. In cells treated with both dexamethasone and geldanamycin, the GFP-GR becomes concentrated in fluorescent globules located periodically along the neurites. Carboxyl terminus of Hsc70-interacting protein (CHIP), the E3 ubiquitin ligase for the GR, also concentrates in the same loci in a steroid-dependent and geldanamycin-dependent manner. If geldanamycin is removed, the GFP-GR exits the globules and continues its retrograde movement. However, in the continued presence of geldanamycin, the GFP-GR in the globules undergoes proteasomal degradation, suggesting that the globules function as degradasomes. This is the first evidence for a linkage between receptor trafficking along neurites and receptor degradation by the proteasome.Fil: Galigniana, Mario Daniel. University of Michigan; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Harrell, Jennifer M.. University of Michigan; Estados UnidosFil: Housley, Paul R.. University of South Carolina; Estados UnidosFil: Patterson, Cam. University of North Carolina; Estados UnidosFil: Fisher, Stephen K.. University of Michigan; Estados UnidosFil: Pratt, William B.. University of Michigan; Estados UnidosElsevier2004-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/29101Galigniana, Mario Daniel; Harrell, Jennifer M.; Housley, Paul R. ; Patterson, Cam ; Fisher, Stephen K. ; et al.; Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome; Elsevier; Molecular Brain Research; 123; 12-2004; 27-360169-328XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0169328X04000269?via%3Dihub#!info:eu-repo/semantics/altIdentifier/doi/10.1016/j.molbrainres.2003.12.015info:eu-repo/semantics/altIdentifier/pmid/15046863info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:03:55Zoai:ri.conicet.gov.ar:11336/29101instacron: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 10:03:55.81CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome
title Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome
spellingShingle Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome
Galigniana, Mario Daniel
Protein Trafficking
Hsp90
Proteasomal Degradation
Glucocorticoid Receptor
Chip
Geldanamycin
Neuronal Cells
title_short Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome
title_full Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome
title_fullStr Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome
title_full_unstemmed Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome
title_sort Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome
dc.creator.none.fl_str_mv Galigniana, Mario Daniel
Harrell, Jennifer M.
Housley, Paul R.
Patterson, Cam
Fisher, Stephen K.
Pratt, William B.
author Galigniana, Mario Daniel
author_facet Galigniana, Mario Daniel
Harrell, Jennifer M.
Housley, Paul R.
Patterson, Cam
Fisher, Stephen K.
Pratt, William B.
author_role author
author2 Harrell, Jennifer M.
Housley, Paul R.
Patterson, Cam
Fisher, Stephen K.
Pratt, William B.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Protein Trafficking
Hsp90
Proteasomal Degradation
Glucocorticoid Receptor
Chip
Geldanamycin
Neuronal Cells
topic Protein Trafficking
Hsp90
Proteasomal Degradation
Glucocorticoid Receptor
Chip
Geldanamycin
Neuronal Cells
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Here, we have used a chimera of green fluorescent protein (GFP) and the glucocorticoid receptor (GR) to study retrograde movement of a model soluble (i.e., non-vesicle-associated) protein in axons and dendrites of cultured NT2-N neurons. It is known that in non-neuronal cells, the GFP-GR moves from cytoplasm to the nucleus in a steroid-dependent manner by a rapid, hsp90-dependent mechanism. When rapid movement is inhibited by geldanamycin (GA), a specific inhibitor of the protein chaperone hsp90, the GFP-GR translocates slowly to the nucleus by diffusion. Here we show that GFP-GR expressed in hormone-free neurons is localized in both cytoplasm and neurites, and upon treatment with dexamethasone (DEX), it moves to the nucleus. In neurites, movement by diffusion is not possible, and we show that movement of the GFP-GR from neurites is blocked by geldanamycin, suggesting that the hsp90-dependent movement machinery is required for retrograde movement. In cells treated with both dexamethasone and geldanamycin, the GFP-GR becomes concentrated in fluorescent globules located periodically along the neurites. Carboxyl terminus of Hsc70-interacting protein (CHIP), the E3 ubiquitin ligase for the GR, also concentrates in the same loci in a steroid-dependent and geldanamycin-dependent manner. If geldanamycin is removed, the GFP-GR exits the globules and continues its retrograde movement. However, in the continued presence of geldanamycin, the GFP-GR in the globules undergoes proteasomal degradation, suggesting that the globules function as degradasomes. This is the first evidence for a linkage between receptor trafficking along neurites and receptor degradation by the proteasome.
Fil: Galigniana, Mario Daniel. University of Michigan; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Harrell, Jennifer M.. University of Michigan; Estados Unidos
Fil: Housley, Paul R.. University of South Carolina; Estados Unidos
Fil: Patterson, Cam. University of North Carolina; Estados Unidos
Fil: Fisher, Stephen K.. University of Michigan; Estados Unidos
Fil: Pratt, William B.. University of Michigan; Estados Unidos
description Here, we have used a chimera of green fluorescent protein (GFP) and the glucocorticoid receptor (GR) to study retrograde movement of a model soluble (i.e., non-vesicle-associated) protein in axons and dendrites of cultured NT2-N neurons. It is known that in non-neuronal cells, the GFP-GR moves from cytoplasm to the nucleus in a steroid-dependent manner by a rapid, hsp90-dependent mechanism. When rapid movement is inhibited by geldanamycin (GA), a specific inhibitor of the protein chaperone hsp90, the GFP-GR translocates slowly to the nucleus by diffusion. Here we show that GFP-GR expressed in hormone-free neurons is localized in both cytoplasm and neurites, and upon treatment with dexamethasone (DEX), it moves to the nucleus. In neurites, movement by diffusion is not possible, and we show that movement of the GFP-GR from neurites is blocked by geldanamycin, suggesting that the hsp90-dependent movement machinery is required for retrograde movement. In cells treated with both dexamethasone and geldanamycin, the GFP-GR becomes concentrated in fluorescent globules located periodically along the neurites. Carboxyl terminus of Hsc70-interacting protein (CHIP), the E3 ubiquitin ligase for the GR, also concentrates in the same loci in a steroid-dependent and geldanamycin-dependent manner. If geldanamycin is removed, the GFP-GR exits the globules and continues its retrograde movement. However, in the continued presence of geldanamycin, the GFP-GR in the globules undergoes proteasomal degradation, suggesting that the globules function as degradasomes. This is the first evidence for a linkage between receptor trafficking along neurites and receptor degradation by the proteasome.
publishDate 2004
dc.date.none.fl_str_mv 2004-12
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/11336/29101
Galigniana, Mario Daniel; Harrell, Jennifer M.; Housley, Paul R. ; Patterson, Cam ; Fisher, Stephen K. ; et al.; Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome; Elsevier; Molecular Brain Research; 123; 12-2004; 27-36
0169-328X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/29101
identifier_str_mv Galigniana, Mario Daniel; Harrell, Jennifer M.; Housley, Paul R. ; Patterson, Cam ; Fisher, Stephen K. ; et al.; Retrograde transport of the GR requires dynamic assembly of complexes with hsp90 and is linked to the CHIP component of the proteosome; Elsevier; Molecular Brain Research; 123; 12-2004; 27-36
0169-328X
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/S0169328X04000269?via%3Dihub#!
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.molbrainres.2003.12.015
info:eu-repo/semantics/altIdentifier/pmid/15046863
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 Elsevier
publisher.none.fl_str_mv Elsevier
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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