Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells

Autores
Tax, Gabor; Guay, Kevin P.; Pantalone, Ludovica; Ceci, Martina; Soldà, Tatiana; Hitchman, Charlie J.; Hill, Johan C.; Vasiljević, Snežana; Lia, Andrea; Modenutti, Carlos Pablo; Straatman, Kees R.; Santino, Angelo; Molinari, Maurizio; Zitzmann, Nicole; Hebert, Daniel N.; Roversi, Pietro; Trerotola, Marco
Año de publicación
2024
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Endoplasmic reticulum (ER) retention of misfolded glycoproteins is mediated by the ER-localized eukaryotic glycoprotein secretion checkpoint, UDP-glucose glycoprotein glucosyl-transferase (UGGT). The enzyme recognizes a misfolded glycoprotein and flags it for ER retention by re-glucosylating one of its N-linked glycans. In the background of a congenital mutation in a secreted glycoprotein gene, UGGT-mediated ER retention can cause rare disease, even if the mutant glycoprotein retains activity (“responsive mutant”). Using confocal laser scanning microscopy, we investigated here the subcellular localization of the human Trop-2-Q118E, E227K and L186P mutants, which cause gelatinous drop-like corneal dystrophy (GDLD). Compared with the wild-type Trop-2, which is correctly localized at the plasma membrane, these Trop-2 mutants are retained in the ER. We studied fluorescent chimeras of the Trop-2 Q118E, E227K and L186P mutants in mammalian cells harboring CRISPR/Cas9-mediated inhibition of the UGGT1 and/or UGGT2 genes. The membrane localization of the Trop-2 Q118E, E227K and L186P mutants was successfully rescued in UGGT1−/−cells. UGGT1 also efficiently reglucosylated Trop-2-Q118E-EYFP in cellula. The study supports the hypothesis that UGGT1 modulation would constitute a novel therapeutic strategy for the treatment of pathological conditions associated to misfolded membrane glycoproteins (whenever the mutation impairs but does not abrogate function), and it encourages the testing of modulators of ER glycoprotein folding quality control as broad-spectrum rescue-of-secretion drugs in rare diseases caused by responsive secreted glycoprotein mutants.
Fil: Tax, Gabor. University of Leicester; Reino Unido
Fil: Guay, Kevin P.. University of Massachusetts; Estados Unidos
Fil: Pantalone, Ludovica. University of Chieti-Pescara; Italia
Fil: Ceci, Martina. University of Chieti-Pescara; Italia
Fil: Soldà, Tatiana. Università della Svizzera Italiana; Italia
Fil: Hitchman, Charlie J.. University of Leicester; Reino Unido
Fil: Hill, Johan C.. University of Oxford; Reino Unido
Fil: Vasiljević, Snežana. University of Oxford; Reino Unido
Fil: Lia, Andrea. University of Leicester; Reino Unido. Consiglio Nazionale delle Ricerche. Istituto di Scienze delle Produzioni Alimentari; Italia
Fil: Modenutti, Carlos Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Straatman, Kees R.. University of Leicester; Reino Unido
Fil: Santino, Angelo. Consiglio Nazionale delle Ricerche. Istituto di Scienze delle Produzioni Alimentari; Italia
Fil: Molinari, Maurizio. Università della Svizzera Italiana; Italia. École Polytechnique Fédérale de Lausanne; Suiza
Fil: Zitzmann, Nicole. University of Oxford; Reino Unido
Fil: Hebert, Daniel N.. University of Massachusetts; Estados Unidos
Fil: Roversi, Pietro. University of Leicester; Reino Unido. Consiglio Nazionale delle Ricerche; Italia
Fil: Trerotola, Marco. University of Chieti-Pescara; Italia
Materia
Glycoprotein secretion
GDLD
Responsive mutant
TACSTD2
Trop-2
UGGT
UGGT1
UGGT2
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:11336/268543
Acceder
id CONICETDig_44091b7a075c0c2e345ce4af782d4db1
oai_identifier_str oai:ri.conicet.gov.ar:11336/268543
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cellsTax, GaborGuay, Kevin P.Pantalone, LudovicaCeci, MartinaSoldà, TatianaHitchman, Charlie J.Hill, Johan C.Vasiljević, SnežanaLia, AndreaModenutti, Carlos PabloStraatman, Kees R.Santino, AngeloMolinari, MaurizioZitzmann, NicoleHebert, Daniel N.Roversi, PietroTrerotola, MarcoGlycoprotein secretionGDLDResponsive mutantTACSTD2Trop-2UGGTUGGT1UGGT2https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Endoplasmic reticulum (ER) retention of misfolded glycoproteins is mediated by the ER-localized eukaryotic glycoprotein secretion checkpoint, UDP-glucose glycoprotein glucosyl-transferase (UGGT). The enzyme recognizes a misfolded glycoprotein and flags it for ER retention by re-glucosylating one of its N-linked glycans. In the background of a congenital mutation in a secreted glycoprotein gene, UGGT-mediated ER retention can cause rare disease, even if the mutant glycoprotein retains activity (“responsive mutant”). Using confocal laser scanning microscopy, we investigated here the subcellular localization of the human Trop-2-Q118E, E227K and L186P mutants, which cause gelatinous drop-like corneal dystrophy (GDLD). Compared with the wild-type Trop-2, which is correctly localized at the plasma membrane, these Trop-2 mutants are retained in the ER. We studied fluorescent chimeras of the Trop-2 Q118E, E227K and L186P mutants in mammalian cells harboring CRISPR/Cas9-mediated inhibition of the UGGT1 and/or UGGT2 genes. The membrane localization of the Trop-2 Q118E, E227K and L186P mutants was successfully rescued in UGGT1−/−cells. UGGT1 also efficiently reglucosylated Trop-2-Q118E-EYFP in cellula. The study supports the hypothesis that UGGT1 modulation would constitute a novel therapeutic strategy for the treatment of pathological conditions associated to misfolded membrane glycoproteins (whenever the mutation impairs but does not abrogate function), and it encourages the testing of modulators of ER glycoprotein folding quality control as broad-spectrum rescue-of-secretion drugs in rare diseases caused by responsive secreted glycoprotein mutants.Fil: Tax, Gabor. University of Leicester; Reino UnidoFil: Guay, Kevin P.. University of Massachusetts; Estados UnidosFil: Pantalone, Ludovica. University of Chieti-Pescara; ItaliaFil: Ceci, Martina. University of Chieti-Pescara; ItaliaFil: Soldà, Tatiana. Università della Svizzera Italiana; ItaliaFil: Hitchman, Charlie J.. University of Leicester; Reino UnidoFil: Hill, Johan C.. University of Oxford; Reino UnidoFil: Vasiljević, Snežana. University of Oxford; Reino UnidoFil: Lia, Andrea. University of Leicester; Reino Unido. Consiglio Nazionale delle Ricerche. Istituto di Scienze delle Produzioni Alimentari; ItaliaFil: Modenutti, Carlos Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Straatman, Kees R.. University of Leicester; Reino UnidoFil: Santino, Angelo. Consiglio Nazionale delle Ricerche. Istituto di Scienze delle Produzioni Alimentari; ItaliaFil: Molinari, Maurizio. Università della Svizzera Italiana; Italia. École Polytechnique Fédérale de Lausanne; SuizaFil: Zitzmann, Nicole. University of Oxford; Reino UnidoFil: Hebert, Daniel N.. University of Massachusetts; Estados UnidosFil: Roversi, Pietro. University of Leicester; Reino Unido. Consiglio Nazionale delle Ricerche; ItaliaFil: Trerotola, Marco. University of Chieti-Pescara; ItaliaWiley Blackwell Publishing, Inc2024-01info: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/11336/268543Tax, Gabor; Guay, Kevin P.; Pantalone, Ludovica; Ceci, Martina; Soldà, Tatiana; et al.; Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells; Wiley Blackwell Publishing, Inc; Traffic; 25; 1; 1-2024; 1-201398-9219CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/10.1111/tra.12927info:eu-repo/semantics/altIdentifier/doi/10.1111/tra.12927info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:44:47Zoai:ri.conicet.gov.ar:11336/268543instacron: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:34982025-10-15 15:44:47.294CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells
title Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells
spellingShingle Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells
Tax, Gabor
Glycoprotein secretion
GDLD
Responsive mutant
TACSTD2
Trop-2
UGGT
UGGT1
UGGT2
title_short Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells
title_full Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells
title_fullStr Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells
title_full_unstemmed Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells
title_sort Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells
dc.creator.none.fl_str_mv Tax, Gabor
Guay, Kevin P.
Pantalone, Ludovica
Ceci, Martina
Soldà, Tatiana
Hitchman, Charlie J.
Hill, Johan C.
Vasiljević, Snežana
Lia, Andrea
Modenutti, Carlos Pablo
Straatman, Kees R.
Santino, Angelo
Molinari, Maurizio
Zitzmann, Nicole
Hebert, Daniel N.
Roversi, Pietro
Trerotola, Marco
author Tax, Gabor
author_facet Tax, Gabor
Guay, Kevin P.
Pantalone, Ludovica
Ceci, Martina
Soldà, Tatiana
Hitchman, Charlie J.
Hill, Johan C.
Vasiljević, Snežana
Lia, Andrea
Modenutti, Carlos Pablo
Straatman, Kees R.
Santino, Angelo
Molinari, Maurizio
Zitzmann, Nicole
Hebert, Daniel N.
Roversi, Pietro
Trerotola, Marco
author_role author
author2 Guay, Kevin P.
Pantalone, Ludovica
Ceci, Martina
Soldà, Tatiana
Hitchman, Charlie J.
Hill, Johan C.
Vasiljević, Snežana
Lia, Andrea
Modenutti, Carlos Pablo
Straatman, Kees R.
Santino, Angelo
Molinari, Maurizio
Zitzmann, Nicole
Hebert, Daniel N.
Roversi, Pietro
Trerotola, Marco
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Glycoprotein secretion
GDLD
Responsive mutant
TACSTD2
Trop-2
UGGT
UGGT1
UGGT2
topic Glycoprotein secretion
GDLD
Responsive mutant
TACSTD2
Trop-2
UGGT
UGGT1
UGGT2
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Endoplasmic reticulum (ER) retention of misfolded glycoproteins is mediated by the ER-localized eukaryotic glycoprotein secretion checkpoint, UDP-glucose glycoprotein glucosyl-transferase (UGGT). The enzyme recognizes a misfolded glycoprotein and flags it for ER retention by re-glucosylating one of its N-linked glycans. In the background of a congenital mutation in a secreted glycoprotein gene, UGGT-mediated ER retention can cause rare disease, even if the mutant glycoprotein retains activity (“responsive mutant”). Using confocal laser scanning microscopy, we investigated here the subcellular localization of the human Trop-2-Q118E, E227K and L186P mutants, which cause gelatinous drop-like corneal dystrophy (GDLD). Compared with the wild-type Trop-2, which is correctly localized at the plasma membrane, these Trop-2 mutants are retained in the ER. We studied fluorescent chimeras of the Trop-2 Q118E, E227K and L186P mutants in mammalian cells harboring CRISPR/Cas9-mediated inhibition of the UGGT1 and/or UGGT2 genes. The membrane localization of the Trop-2 Q118E, E227K and L186P mutants was successfully rescued in UGGT1−/−cells. UGGT1 also efficiently reglucosylated Trop-2-Q118E-EYFP in cellula. The study supports the hypothesis that UGGT1 modulation would constitute a novel therapeutic strategy for the treatment of pathological conditions associated to misfolded membrane glycoproteins (whenever the mutation impairs but does not abrogate function), and it encourages the testing of modulators of ER glycoprotein folding quality control as broad-spectrum rescue-of-secretion drugs in rare diseases caused by responsive secreted glycoprotein mutants.
Fil: Tax, Gabor. University of Leicester; Reino Unido
Fil: Guay, Kevin P.. University of Massachusetts; Estados Unidos
Fil: Pantalone, Ludovica. University of Chieti-Pescara; Italia
Fil: Ceci, Martina. University of Chieti-Pescara; Italia
Fil: Soldà, Tatiana. Università della Svizzera Italiana; Italia
Fil: Hitchman, Charlie J.. University of Leicester; Reino Unido
Fil: Hill, Johan C.. University of Oxford; Reino Unido
Fil: Vasiljević, Snežana. University of Oxford; Reino Unido
Fil: Lia, Andrea. University of Leicester; Reino Unido. Consiglio Nazionale delle Ricerche. Istituto di Scienze delle Produzioni Alimentari; Italia
Fil: Modenutti, Carlos Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Straatman, Kees R.. University of Leicester; Reino Unido
Fil: Santino, Angelo. Consiglio Nazionale delle Ricerche. Istituto di Scienze delle Produzioni Alimentari; Italia
Fil: Molinari, Maurizio. Università della Svizzera Italiana; Italia. École Polytechnique Fédérale de Lausanne; Suiza
Fil: Zitzmann, Nicole. University of Oxford; Reino Unido
Fil: Hebert, Daniel N.. University of Massachusetts; Estados Unidos
Fil: Roversi, Pietro. University of Leicester; Reino Unido. Consiglio Nazionale delle Ricerche; Italia
Fil: Trerotola, Marco. University of Chieti-Pescara; Italia
description Endoplasmic reticulum (ER) retention of misfolded glycoproteins is mediated by the ER-localized eukaryotic glycoprotein secretion checkpoint, UDP-glucose glycoprotein glucosyl-transferase (UGGT). The enzyme recognizes a misfolded glycoprotein and flags it for ER retention by re-glucosylating one of its N-linked glycans. In the background of a congenital mutation in a secreted glycoprotein gene, UGGT-mediated ER retention can cause rare disease, even if the mutant glycoprotein retains activity (“responsive mutant”). Using confocal laser scanning microscopy, we investigated here the subcellular localization of the human Trop-2-Q118E, E227K and L186P mutants, which cause gelatinous drop-like corneal dystrophy (GDLD). Compared with the wild-type Trop-2, which is correctly localized at the plasma membrane, these Trop-2 mutants are retained in the ER. We studied fluorescent chimeras of the Trop-2 Q118E, E227K and L186P mutants in mammalian cells harboring CRISPR/Cas9-mediated inhibition of the UGGT1 and/or UGGT2 genes. The membrane localization of the Trop-2 Q118E, E227K and L186P mutants was successfully rescued in UGGT1−/−cells. UGGT1 also efficiently reglucosylated Trop-2-Q118E-EYFP in cellula. The study supports the hypothesis that UGGT1 modulation would constitute a novel therapeutic strategy for the treatment of pathological conditions associated to misfolded membrane glycoproteins (whenever the mutation impairs but does not abrogate function), and it encourages the testing of modulators of ER glycoprotein folding quality control as broad-spectrum rescue-of-secretion drugs in rare diseases caused by responsive secreted glycoprotein mutants.
publishDate 2024
dc.date.none.fl_str_mv 2024-01
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/11336/268543
Tax, Gabor; Guay, Kevin P.; Pantalone, Ludovica; Ceci, Martina; Soldà, Tatiana; et al.; Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells; Wiley Blackwell Publishing, Inc; Traffic; 25; 1; 1-2024; 1-20
1398-9219
CONICET Digital
CONICET
url http://hdl.handle.net/11336/268543
identifier_str_mv Tax, Gabor; Guay, Kevin P.; Pantalone, Ludovica; Ceci, Martina; Soldà, Tatiana; et al.; Rescue of secretion of rare‐disease‐associated misfolded mutant glycoproteins in UGGT1 knock‐out mammalian cells; Wiley Blackwell Publishing, Inc; Traffic; 25; 1; 1-2024; 1-20
1398-9219
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://onlinelibrary.wiley.com/doi/10.1111/tra.12927
info:eu-repo/semantics/altIdentifier/doi/10.1111/tra.12927
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 Wiley Blackwell Publishing, Inc
publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
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 12.891075

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