GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis

Autores
Zibrova, Darya; Vandermoere, Franck; Göransson, Olga; Peggie, Mark; Mariño, Karina Valeria; Knierim, Anne; Spengler, Katrin; Weigert, Cora; Viollet, Benoit; Morrice, Nicholas A.; Sakamoto, Kei; Heller, Regine
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Activation of AMP-activated protein kinase (AMPK) in endothelial cells regulates energy homeostasis, stress protection and angiogenesis, but the underlying mechanisms are incompletely understood. Using a label-free phosphoproteomic analysis, we identified glutamine:fructose-6-phosphate amidotransferase 1 (GFAT1) as an AMPK substrate. GFAT1 is the rate-limiting enzyme in the hexosamine biosynthesis pathway (HBP) and as such controls the modification of proteins by O-linked β-N-acetylglucosamine (O-GlcNAc). In the present study, we tested the hypothesis that AMPK controls O-GlcNAc levels and function of endothelial cells via GFAT1 phosphorylation using biochemical, pharmacological, genetic and in vitro angiogenesis approaches. Activation of AMPK in primary human endothelial cells by 5-aminoimidazole-4-carboxamide riboside (AICAR) or by vascular endothelial growth factor (VEGF) led to GFAT1 phosphorylation at serine 243. This effect was not seen when AMPK was down-regulated by siRNA. Upon AMPK activation, diminished GFAT activity and reduced O-GlcNAc levels were observed in endothelial cells containing wild-type (WT)-GFAT1 but not in cells expressing non-phosphorylatable S243A-GFAT1. Pharmacological inhibition or siRNA-mediated down-regulation of GFAT1 potentiated VEGF-induced sprouting, indicating that GFAT1 acts as a negative regulator of angiogenesis. In cells expressing S243A-GFAT1, VEGF-induced sprouting was reduced, suggesting that VEGF relieves the inhibitory action of GFAT1/HBP on angiogenesis via AMPK-mediated GFAT1 phosphorylation. Activation of GFAT1/HBP by high glucose led to impairment of vascular sprouting, whereas GFAT1 inhibition improved sprouting even if glucose level was high. Our findings provide novel mechanistic insights into the role of HBP in angiogenesis. They suggest that targeting AMPK in endothelium might help to ameliorate hyperglycaemia-induced vascular dysfunction associated with metabolic disorders.
Fil: Zibrova, Darya. Universitat Jena; Alemania
Fil: Vandermoere, Franck. Universite de Montpellier; Francia. Inserm; Francia
Fil: Göransson, Olga. Lund University; Suecia
Fil: Peggie, Mark. University of Dundee; Reino Unido
Fil: Mariño, Karina Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Knierim, Anne. Universitat Jena; Alemania
Fil: Spengler, Katrin. Universitat Jena; Alemania
Fil: Weigert, Cora. German Center For Diabetes Research ; Alemania. Universitat Tübingen; Alemania
Fil: Viollet, Benoit. Inserm; Francia. Université Paris Descartes; Francia
Fil: Morrice, Nicholas A.. AB-Sciex; Reino Unido
Fil: Sakamoto, Kei. University Of Dundee;
Fil: Heller, Regine. Universitat Jena; Alemania
Materia
Ampk
Angiogenesis
Gfat1
O-Glcnacylation
Phosphoproteomics
Vegf
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/24568
Acceder
id CONICETDig_3fe59eb02186a82d770592e7a8c343f5
oai_identifier_str oai:ri.conicet.gov.ar:11336/24568
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesisZibrova, DaryaVandermoere, FranckGöransson, OlgaPeggie, MarkMariño, Karina ValeriaKnierim, AnneSpengler, KatrinWeigert, CoraViollet, BenoitMorrice, Nicholas A.Sakamoto, KeiHeller, RegineAmpkAngiogenesisGfat1O-GlcnacylationPhosphoproteomicsVegfhttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Activation of AMP-activated protein kinase (AMPK) in endothelial cells regulates energy homeostasis, stress protection and angiogenesis, but the underlying mechanisms are incompletely understood. Using a label-free phosphoproteomic analysis, we identified glutamine:fructose-6-phosphate amidotransferase 1 (GFAT1) as an AMPK substrate. GFAT1 is the rate-limiting enzyme in the hexosamine biosynthesis pathway (HBP) and as such controls the modification of proteins by O-linked β-N-acetylglucosamine (O-GlcNAc). In the present study, we tested the hypothesis that AMPK controls O-GlcNAc levels and function of endothelial cells via GFAT1 phosphorylation using biochemical, pharmacological, genetic and in vitro angiogenesis approaches. Activation of AMPK in primary human endothelial cells by 5-aminoimidazole-4-carboxamide riboside (AICAR) or by vascular endothelial growth factor (VEGF) led to GFAT1 phosphorylation at serine 243. This effect was not seen when AMPK was down-regulated by siRNA. Upon AMPK activation, diminished GFAT activity and reduced O-GlcNAc levels were observed in endothelial cells containing wild-type (WT)-GFAT1 but not in cells expressing non-phosphorylatable S243A-GFAT1. Pharmacological inhibition or siRNA-mediated down-regulation of GFAT1 potentiated VEGF-induced sprouting, indicating that GFAT1 acts as a negative regulator of angiogenesis. In cells expressing S243A-GFAT1, VEGF-induced sprouting was reduced, suggesting that VEGF relieves the inhibitory action of GFAT1/HBP on angiogenesis via AMPK-mediated GFAT1 phosphorylation. Activation of GFAT1/HBP by high glucose led to impairment of vascular sprouting, whereas GFAT1 inhibition improved sprouting even if glucose level was high. Our findings provide novel mechanistic insights into the role of HBP in angiogenesis. They suggest that targeting AMPK in endothelium might help to ameliorate hyperglycaemia-induced vascular dysfunction associated with metabolic disorders.Fil: Zibrova, Darya. Universitat Jena; AlemaniaFil: Vandermoere, Franck. Universite de Montpellier; Francia. Inserm; FranciaFil: Göransson, Olga. Lund University; SueciaFil: Peggie, Mark. University of Dundee; Reino UnidoFil: Mariño, Karina Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Knierim, Anne. Universitat Jena; AlemaniaFil: Spengler, Katrin. Universitat Jena; AlemaniaFil: Weigert, Cora. German Center For Diabetes Research ; Alemania. Universitat Tübingen; AlemaniaFil: Viollet, Benoit. Inserm; Francia. Université Paris Descartes; FranciaFil: Morrice, Nicholas A.. AB-Sciex; Reino UnidoFil: Sakamoto, Kei. University Of Dundee;Fil: Heller, Regine. Universitat Jena; AlemaniaPortland Press2017-03-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/24568Zibrova, Darya; Vandermoere, Franck; Göransson, Olga; Peggie, Mark; Mariño, Karina Valeria; et al.; GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis; Portland Press; Biochemical Journal; 474; 6; 7-3-2017; 983-10010264-60211470-8728CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/info:eu-repo/semantics/altIdentifier/doi/10.1042/BCJ20160980info:eu-repo/semantics/altIdentifier/pmid/28008135info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:01:02Zoai:ri.conicet.gov.ar:11336/24568instacron: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 10:01:02.88CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis
title GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis
spellingShingle GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis
Zibrova, Darya
Ampk
Angiogenesis
Gfat1
O-Glcnacylation
Phosphoproteomics
Vegf
title_short GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis
title_full GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis
title_fullStr GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis
title_full_unstemmed GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis
title_sort GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis
dc.creator.none.fl_str_mv Zibrova, Darya
Vandermoere, Franck
Göransson, Olga
Peggie, Mark
Mariño, Karina Valeria
Knierim, Anne
Spengler, Katrin
Weigert, Cora
Viollet, Benoit
Morrice, Nicholas A.
Sakamoto, Kei
Heller, Regine
author Zibrova, Darya
author_facet Zibrova, Darya
Vandermoere, Franck
Göransson, Olga
Peggie, Mark
Mariño, Karina Valeria
Knierim, Anne
Spengler, Katrin
Weigert, Cora
Viollet, Benoit
Morrice, Nicholas A.
Sakamoto, Kei
Heller, Regine
author_role author
author2 Vandermoere, Franck
Göransson, Olga
Peggie, Mark
Mariño, Karina Valeria
Knierim, Anne
Spengler, Katrin
Weigert, Cora
Viollet, Benoit
Morrice, Nicholas A.
Sakamoto, Kei
Heller, Regine
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Ampk
Angiogenesis
Gfat1
O-Glcnacylation
Phosphoproteomics
Vegf
topic Ampk
Angiogenesis
Gfat1
O-Glcnacylation
Phosphoproteomics
Vegf
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Activation of AMP-activated protein kinase (AMPK) in endothelial cells regulates energy homeostasis, stress protection and angiogenesis, but the underlying mechanisms are incompletely understood. Using a label-free phosphoproteomic analysis, we identified glutamine:fructose-6-phosphate amidotransferase 1 (GFAT1) as an AMPK substrate. GFAT1 is the rate-limiting enzyme in the hexosamine biosynthesis pathway (HBP) and as such controls the modification of proteins by O-linked β-N-acetylglucosamine (O-GlcNAc). In the present study, we tested the hypothesis that AMPK controls O-GlcNAc levels and function of endothelial cells via GFAT1 phosphorylation using biochemical, pharmacological, genetic and in vitro angiogenesis approaches. Activation of AMPK in primary human endothelial cells by 5-aminoimidazole-4-carboxamide riboside (AICAR) or by vascular endothelial growth factor (VEGF) led to GFAT1 phosphorylation at serine 243. This effect was not seen when AMPK was down-regulated by siRNA. Upon AMPK activation, diminished GFAT activity and reduced O-GlcNAc levels were observed in endothelial cells containing wild-type (WT)-GFAT1 but not in cells expressing non-phosphorylatable S243A-GFAT1. Pharmacological inhibition or siRNA-mediated down-regulation of GFAT1 potentiated VEGF-induced sprouting, indicating that GFAT1 acts as a negative regulator of angiogenesis. In cells expressing S243A-GFAT1, VEGF-induced sprouting was reduced, suggesting that VEGF relieves the inhibitory action of GFAT1/HBP on angiogenesis via AMPK-mediated GFAT1 phosphorylation. Activation of GFAT1/HBP by high glucose led to impairment of vascular sprouting, whereas GFAT1 inhibition improved sprouting even if glucose level was high. Our findings provide novel mechanistic insights into the role of HBP in angiogenesis. They suggest that targeting AMPK in endothelium might help to ameliorate hyperglycaemia-induced vascular dysfunction associated with metabolic disorders.
Fil: Zibrova, Darya. Universitat Jena; Alemania
Fil: Vandermoere, Franck. Universite de Montpellier; Francia. Inserm; Francia
Fil: Göransson, Olga. Lund University; Suecia
Fil: Peggie, Mark. University of Dundee; Reino Unido
Fil: Mariño, Karina Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Knierim, Anne. Universitat Jena; Alemania
Fil: Spengler, Katrin. Universitat Jena; Alemania
Fil: Weigert, Cora. German Center For Diabetes Research ; Alemania. Universitat Tübingen; Alemania
Fil: Viollet, Benoit. Inserm; Francia. Université Paris Descartes; Francia
Fil: Morrice, Nicholas A.. AB-Sciex; Reino Unido
Fil: Sakamoto, Kei. University Of Dundee;
Fil: Heller, Regine. Universitat Jena; Alemania
description Activation of AMP-activated protein kinase (AMPK) in endothelial cells regulates energy homeostasis, stress protection and angiogenesis, but the underlying mechanisms are incompletely understood. Using a label-free phosphoproteomic analysis, we identified glutamine:fructose-6-phosphate amidotransferase 1 (GFAT1) as an AMPK substrate. GFAT1 is the rate-limiting enzyme in the hexosamine biosynthesis pathway (HBP) and as such controls the modification of proteins by O-linked β-N-acetylglucosamine (O-GlcNAc). In the present study, we tested the hypothesis that AMPK controls O-GlcNAc levels and function of endothelial cells via GFAT1 phosphorylation using biochemical, pharmacological, genetic and in vitro angiogenesis approaches. Activation of AMPK in primary human endothelial cells by 5-aminoimidazole-4-carboxamide riboside (AICAR) or by vascular endothelial growth factor (VEGF) led to GFAT1 phosphorylation at serine 243. This effect was not seen when AMPK was down-regulated by siRNA. Upon AMPK activation, diminished GFAT activity and reduced O-GlcNAc levels were observed in endothelial cells containing wild-type (WT)-GFAT1 but not in cells expressing non-phosphorylatable S243A-GFAT1. Pharmacological inhibition or siRNA-mediated down-regulation of GFAT1 potentiated VEGF-induced sprouting, indicating that GFAT1 acts as a negative regulator of angiogenesis. In cells expressing S243A-GFAT1, VEGF-induced sprouting was reduced, suggesting that VEGF relieves the inhibitory action of GFAT1/HBP on angiogenesis via AMPK-mediated GFAT1 phosphorylation. Activation of GFAT1/HBP by high glucose led to impairment of vascular sprouting, whereas GFAT1 inhibition improved sprouting even if glucose level was high. Our findings provide novel mechanistic insights into the role of HBP in angiogenesis. They suggest that targeting AMPK in endothelium might help to ameliorate hyperglycaemia-induced vascular dysfunction associated with metabolic disorders.
publishDate 2017
dc.date.none.fl_str_mv 2017-03-07
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/11336/24568
Zibrova, Darya; Vandermoere, Franck; Göransson, Olga; Peggie, Mark; Mariño, Karina Valeria; et al.; GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis; Portland Press; Biochemical Journal; 474; 6; 7-3-2017; 983-1001
0264-6021
1470-8728
CONICET Digital
CONICET
url http://hdl.handle.net/11336/24568
identifier_str_mv Zibrova, Darya; Vandermoere, Franck; Göransson, Olga; Peggie, Mark; Mariño, Karina Valeria; et al.; GFAT1 phosphorylation by AMPK promotes VEGF-induced angiogenesis; Portland Press; Biochemical Journal; 474; 6; 7-3-2017; 983-1001
0264-6021
1470-8728
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/
info:eu-repo/semantics/altIdentifier/doi/10.1042/BCJ20160980
info:eu-repo/semantics/altIdentifier/pmid/28008135
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Portland Press
publisher.none.fl_str_mv Portland 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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