Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi...

Autores
Sternlieb, Tamara; Schoijet, Alejandra Cecilia; Genta, Patricio D.; Alonso, Guillermo Daniel
Año de publicación
2019
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The AMP-activated protein kinase (AMPK) is a heterotrimeric enzyme involved in maintaining energy homeostasis in response to different stresses in many organisms. During the transition between the mammalian host and the insect vector, Trypanosoma cruzi, the causative agent of Chagas disease, faces different types of environmental fluctuations, all of which prompt the parasite to remodel its metabolism. Recently, it was shown that Trypanosoma brucei AMPK is involved in the differentiation from the bloodstream slender to stumpy stage and in surface protein expression changes in response to nutritional stress. This underscores the relevance of AMPK for parasite life cycle progression.We identified four candidate genes for the AMPK subunits of T. cruzi (alpha1 and alpha2 catalytic subunits, beta and gamma regulatory subunits). The beta and gamma subunits are largely conserved in their domain structure relative to the mammalian orthologs. However, the alpha subunits show significant sequence and structure differences from the human counterparts. The presence of these subunits in T. cruzi epimastigotes was confirmed by RT-PCR, Western blot using a phospho-AMPKa specific antibody, mass spectrometry and by kinase activity assays using the specific AMPK substrate SAMS. TcAMPKa1 over-expressing epimastigotes showed a lower growth rate in basal culture conditions compared to the control. On the other hand, alpha2 over-expression had the opposite effect. Additionally, we observed upregulation of AMPK activity under epimastigote starvation, and that dorsomorphin, a specific AMPK inhibitor, also inhibits T. cruzi AMPK. Moreover, each of these subunits could complement S. cerevisiae conditional mutants lacking the respective subunit of the AMPK ortholog SNF1. Finally, starving assays with AMPKa over-expressing parasites also showed a possible role of AMPK in autophagy. Overall, our results show for the first time, the presence of a functional AMPK orthologue in Trypanosoma cruzi.
Fil: Sternlieb, Tamara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Schoijet, Alejandra Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Genta, Patricio D.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Alonso, Guillermo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
LXIV Annual Meeting of Sociedad Argentina de Investigación Clínica (SAIC) ;LI Annual Meeting of Asociación Argentina de Farmacología Experimental (SAFE); XXI Annual Meeting of Sociedad Argentina de Biología (SAB); XXXI Annual Meeting of Sociedad Argentina de Protozoología (SAP); IX Annual Meeting of Asociación Argentina de Nanomedicinas (NANOMED-ar); VI Regional Scientific Meeting of Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio (AACyTAL)
Mar del Plata
Argentina
Sociedad Argentina de Protozoología
Sociedad Argentina de Investigación Clínica
Asociación Argentina de Farmacología Experimenta
Sociedad Argentina de Biología
Asociación Argentina de Nanomedicinas
Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Materia
ENERGY METABOLISM
AUTOPHAGY
KINASE
CARBON SOURCE
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:11336/153727
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/153727
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruziSternlieb, TamaraSchoijet, Alejandra CeciliaGenta, Patricio D.Alonso, Guillermo DanielENERGY METABOLISMAUTOPHAGYKINASECARBON SOURCEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The AMP-activated protein kinase (AMPK) is a heterotrimeric enzyme involved in maintaining energy homeostasis in response to different stresses in many organisms. During the transition between the mammalian host and the insect vector, Trypanosoma cruzi, the causative agent of Chagas disease, faces different types of environmental fluctuations, all of which prompt the parasite to remodel its metabolism. Recently, it was shown that Trypanosoma brucei AMPK is involved in the differentiation from the bloodstream slender to stumpy stage and in surface protein expression changes in response to nutritional stress. This underscores the relevance of AMPK for parasite life cycle progression.We identified four candidate genes for the AMPK subunits of T. cruzi (alpha1 and alpha2 catalytic subunits, beta and gamma regulatory subunits). The beta and gamma subunits are largely conserved in their domain structure relative to the mammalian orthologs. However, the alpha subunits show significant sequence and structure differences from the human counterparts. The presence of these subunits in T. cruzi epimastigotes was confirmed by RT-PCR, Western blot using a phospho-AMPKa specific antibody, mass spectrometry and by kinase activity assays using the specific AMPK substrate SAMS. TcAMPKa1 over-expressing epimastigotes showed a lower growth rate in basal culture conditions compared to the control. On the other hand, alpha2 over-expression had the opposite effect. Additionally, we observed upregulation of AMPK activity under epimastigote starvation, and that dorsomorphin, a specific AMPK inhibitor, also inhibits T. cruzi AMPK. Moreover, each of these subunits could complement S. cerevisiae conditional mutants lacking the respective subunit of the AMPK ortholog SNF1. Finally, starving assays with AMPKa over-expressing parasites also showed a possible role of AMPK in autophagy. Overall, our results show for the first time, the presence of a functional AMPK orthologue in Trypanosoma cruzi.Fil: Sternlieb, Tamara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Schoijet, Alejandra Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Genta, Patricio D.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Alonso, Guillermo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaLXIV Annual Meeting of Sociedad Argentina de Investigación Clínica (SAIC) ;LI Annual Meeting of Asociación Argentina de Farmacología Experimental (SAFE); XXI Annual Meeting of Sociedad Argentina de Biología (SAB); XXXI Annual Meeting of Sociedad Argentina de Protozoología (SAP); IX Annual Meeting of Asociación Argentina de Nanomedicinas (NANOMED-ar); VI Regional Scientific Meeting of Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio (AACyTAL)Mar del PlataArgentinaSociedad Argentina de ProtozoologíaSociedad Argentina de Investigación ClínicaAsociación Argentina de Farmacología ExperimentaSociedad Argentina de BiologíaAsociación Argentina de NanomedicinasAsociación Argentina de Ciencia y Tecnología de Animales de LaboratorioFundación Revista Medicina2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/153727Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi; LXIV Annual Meeting of Sociedad Argentina de Investigación Clínica (SAIC) ;LI Annual Meeting of Asociación Argentina de Farmacología Experimental (SAFE); XXI Annual Meeting of Sociedad Argentina de Biología (SAB); XXXI Annual Meeting of Sociedad Argentina de Protozoología (SAP); IX Annual Meeting of Asociación Argentina de Nanomedicinas (NANOMED-ar); VI Regional Scientific Meeting of Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio (AACyTAL); Mar del Plata; Argentina; 2019; 1-40025-7680CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.medicinabuenosaires.com/PMID/32692316.pdfInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-17T11:02:09Zoai:ri.conicet.gov.ar:11336/153727instacron: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:34982025-09-17 11:02:10.187CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi
title Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi
spellingShingle Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi
Sternlieb, Tamara
ENERGY METABOLISM
AUTOPHAGY
KINASE
CARBON SOURCE
title_short Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi
title_full Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi
title_fullStr Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi
title_full_unstemmed Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi
title_sort Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi
dc.creator.none.fl_str_mv Sternlieb, Tamara
Schoijet, Alejandra Cecilia
Genta, Patricio D.
Alonso, Guillermo Daniel
author Sternlieb, Tamara
author_facet Sternlieb, Tamara
Schoijet, Alejandra Cecilia
Genta, Patricio D.
Alonso, Guillermo Daniel
author_role author
author2 Schoijet, Alejandra Cecilia
Genta, Patricio D.
Alonso, Guillermo Daniel
author2_role author
author
author
dc.subject.none.fl_str_mv ENERGY METABOLISM
AUTOPHAGY
KINASE
CARBON SOURCE
topic ENERGY METABOLISM
AUTOPHAGY
KINASE
CARBON SOURCE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The AMP-activated protein kinase (AMPK) is a heterotrimeric enzyme involved in maintaining energy homeostasis in response to different stresses in many organisms. During the transition between the mammalian host and the insect vector, Trypanosoma cruzi, the causative agent of Chagas disease, faces different types of environmental fluctuations, all of which prompt the parasite to remodel its metabolism. Recently, it was shown that Trypanosoma brucei AMPK is involved in the differentiation from the bloodstream slender to stumpy stage and in surface protein expression changes in response to nutritional stress. This underscores the relevance of AMPK for parasite life cycle progression.We identified four candidate genes for the AMPK subunits of T. cruzi (alpha1 and alpha2 catalytic subunits, beta and gamma regulatory subunits). The beta and gamma subunits are largely conserved in their domain structure relative to the mammalian orthologs. However, the alpha subunits show significant sequence and structure differences from the human counterparts. The presence of these subunits in T. cruzi epimastigotes was confirmed by RT-PCR, Western blot using a phospho-AMPKa specific antibody, mass spectrometry and by kinase activity assays using the specific AMPK substrate SAMS. TcAMPKa1 over-expressing epimastigotes showed a lower growth rate in basal culture conditions compared to the control. On the other hand, alpha2 over-expression had the opposite effect. Additionally, we observed upregulation of AMPK activity under epimastigote starvation, and that dorsomorphin, a specific AMPK inhibitor, also inhibits T. cruzi AMPK. Moreover, each of these subunits could complement S. cerevisiae conditional mutants lacking the respective subunit of the AMPK ortholog SNF1. Finally, starving assays with AMPKa over-expressing parasites also showed a possible role of AMPK in autophagy. Overall, our results show for the first time, the presence of a functional AMPK orthologue in Trypanosoma cruzi.
Fil: Sternlieb, Tamara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Schoijet, Alejandra Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Genta, Patricio D.. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Alonso, Guillermo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
LXIV Annual Meeting of Sociedad Argentina de Investigación Clínica (SAIC) ;LI Annual Meeting of Asociación Argentina de Farmacología Experimental (SAFE); XXI Annual Meeting of Sociedad Argentina de Biología (SAB); XXXI Annual Meeting of Sociedad Argentina de Protozoología (SAP); IX Annual Meeting of Asociación Argentina de Nanomedicinas (NANOMED-ar); VI Regional Scientific Meeting of Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio (AACyTAL)
Mar del Plata
Argentina
Sociedad Argentina de Protozoología
Sociedad Argentina de Investigación Clínica
Asociación Argentina de Farmacología Experimenta
Sociedad Argentina de Biología
Asociación Argentina de Nanomedicinas
Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
description The AMP-activated protein kinase (AMPK) is a heterotrimeric enzyme involved in maintaining energy homeostasis in response to different stresses in many organisms. During the transition between the mammalian host and the insect vector, Trypanosoma cruzi, the causative agent of Chagas disease, faces different types of environmental fluctuations, all of which prompt the parasite to remodel its metabolism. Recently, it was shown that Trypanosoma brucei AMPK is involved in the differentiation from the bloodstream slender to stumpy stage and in surface protein expression changes in response to nutritional stress. This underscores the relevance of AMPK for parasite life cycle progression.We identified four candidate genes for the AMPK subunits of T. cruzi (alpha1 and alpha2 catalytic subunits, beta and gamma regulatory subunits). The beta and gamma subunits are largely conserved in their domain structure relative to the mammalian orthologs. However, the alpha subunits show significant sequence and structure differences from the human counterparts. The presence of these subunits in T. cruzi epimastigotes was confirmed by RT-PCR, Western blot using a phospho-AMPKa specific antibody, mass spectrometry and by kinase activity assays using the specific AMPK substrate SAMS. TcAMPKa1 over-expressing epimastigotes showed a lower growth rate in basal culture conditions compared to the control. On the other hand, alpha2 over-expression had the opposite effect. Additionally, we observed upregulation of AMPK activity under epimastigote starvation, and that dorsomorphin, a specific AMPK inhibitor, also inhibits T. cruzi AMPK. Moreover, each of these subunits could complement S. cerevisiae conditional mutants lacking the respective subunit of the AMPK ortholog SNF1. Finally, starving assays with AMPKa over-expressing parasites also showed a possible role of AMPK in autophagy. Overall, our results show for the first time, the presence of a functional AMPK orthologue in Trypanosoma cruzi.
publishDate 2019
dc.date.none.fl_str_mv 2019
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/153727
Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi; LXIV Annual Meeting of Sociedad Argentina de Investigación Clínica (SAIC) ;LI Annual Meeting of Asociación Argentina de Farmacología Experimental (SAFE); XXI Annual Meeting of Sociedad Argentina de Biología (SAB); XXXI Annual Meeting of Sociedad Argentina de Protozoología (SAP); IX Annual Meeting of Asociación Argentina de Nanomedicinas (NANOMED-ar); VI Regional Scientific Meeting of Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio (AACyTAL); Mar del Plata; Argentina; 2019; 1-4
0025-7680
CONICET Digital
CONICET
url http://hdl.handle.net/11336/153727
identifier_str_mv Functional roles of AMP-activated protein kinase (AMPK) complexes containing TcAMPKα1 or TcAMPKα2 in energy homeostasis regulation and cell culture progression in Trypanosoma cruzi; LXIV Annual Meeting of Sociedad Argentina de Investigación Clínica (SAIC) ;LI Annual Meeting of Asociación Argentina de Farmacología Experimental (SAFE); XXI Annual Meeting of Sociedad Argentina de Biología (SAB); XXXI Annual Meeting of Sociedad Argentina de Protozoología (SAP); IX Annual Meeting of Asociación Argentina de Nanomedicinas (NANOMED-ar); VI Regional Scientific Meeting of Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio (AACyTAL); Mar del Plata; Argentina; 2019; 1-4
0025-7680
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.medicinabuenosaires.com/PMID/32692316.pdf
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
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application/pdf
application/pdf
dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
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
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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