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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/29101
Ver los metadatos del registro completo
id |
CONICETDig_5feb3f84092f5ab9177d9d0d806f60c6 |
---|---|
oai_identifier_str |
oai:ri.conicet.gov.ar:11336/29101 |
network_acronym_str |
CONICETDig |
repository_id_str |
3498 |
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 |
_version_ |
1844613861130371072 |
score |
13.070432 |