A novel approach to study epigenetic marks in Trypanosoma cruzi

Autores
López, María del Rosario; Lavignolle, María del Rosario; Vilchez Larrea, Salomé Catalina; Ocampo, Josefina; Argüello, Rafael; Alonso, Guillermo Daniel
Año de publicación
2023
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
T. cruzi is the etiologic agent of Chagas disease and has a complex life cycle. Epimastigotes are the replicativeform in the vector, trypomastigotes are the infective stage, and amastigotes are a replicative intracellular stage.Proper cell cycle progression requires multiple factors.Epigenetics could be relevant for gene regulation. Particularly, lysine 76 of histone 3 (H3K76), could be mono,di or tri methylated by the methyltransferases TcDot1a and TcDot1b. It is not known how H3K76 methylationinfluences cell cycle progression, but deletion of TcDOT1b arrests the cell cycle in G2/M. Additionally,overexpression of Aurora kinase 1 (TcAUK1), affects the same stage.One of the main problems to study the epigenetic role of H3K76 methylation, or TcAUK1 in cell cycle progressionis that transgenic parasites for these proteins had been hard to obtain or maintain. Therefore, we used Flowcytometry that allowed us to identify the differential methylation of H3K76 and to make quantitative analysisusing low amounts of antibodies. Remarkably, we have been able to fine-tune the technique to this end. Wehave used three strain of T. cruzi, Dm28c; K98 and Tulahuen that differ by their virulence. We could detect byFlow cytometer that H3K76 could be mono, di and trimethylated, being the last one the most abundant mark inevery strain tested. This result is consistent with previous outcomes that showed that H3K76me3 is present inevery step of the cell cycle. Additionally, we could detect TcAUK1 in Dm28c, K98 and Tulahuen. This isimportant because the subcellular location and expression levels of this protein are finely regulated and it isevasive to Western blot detection.Furthermore, to check the potential connection between TcAUK1 and TcDOT1 activity, we are testing differentialH3K76 methylation in parasites that overexpress TcAUK1.Our findings surpass our study, since this approach will be useful for the whole community working intrypanosome epigenetics.
Fil: López, María del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Lavignolle, María del Rosario. Centre D'immunologie Marseille-Luminy; Francia
Fil: Vilchez Larrea, Salomé Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Ocampo, Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Argüello, Rafael. Centre D'immunologie Marseille-Luminy; Francia
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
XXXVIII Annual Meeting of the Brazilian Society of Protozoology; XLIX Annual Meeting on Basic Research in Chagas’ Disease
Caxambú
Brasil
Brazilian Society of Protozoology
Materia
TRYPANOSOMATIDS
FLOW CITOMETRY
HISTONE METHYL TRANSFERASES
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:11336/211751
Acceder
id CONICETDig_b0da635f701040078601abde794682da
oai_identifier_str oai:ri.conicet.gov.ar:11336/211751
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A novel approach to study epigenetic marks in Trypanosoma cruziLópez, María del RosarioLavignolle, María del RosarioVilchez Larrea, Salomé CatalinaOcampo, JosefinaArgüello, RafaelAlonso, Guillermo DanielTRYPANOSOMATIDSFLOW CITOMETRYHISTONE METHYL TRANSFERASEShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1T. cruzi is the etiologic agent of Chagas disease and has a complex life cycle. Epimastigotes are the replicativeform in the vector, trypomastigotes are the infective stage, and amastigotes are a replicative intracellular stage.Proper cell cycle progression requires multiple factors.Epigenetics could be relevant for gene regulation. Particularly, lysine 76 of histone 3 (H3K76), could be mono,di or tri methylated by the methyltransferases TcDot1a and TcDot1b. It is not known how H3K76 methylationinfluences cell cycle progression, but deletion of TcDOT1b arrests the cell cycle in G2/M. Additionally,overexpression of Aurora kinase 1 (TcAUK1), affects the same stage.One of the main problems to study the epigenetic role of H3K76 methylation, or TcAUK1 in cell cycle progressionis that transgenic parasites for these proteins had been hard to obtain or maintain. Therefore, we used Flowcytometry that allowed us to identify the differential methylation of H3K76 and to make quantitative analysisusing low amounts of antibodies. Remarkably, we have been able to fine-tune the technique to this end. Wehave used three strain of T. cruzi, Dm28c; K98 and Tulahuen that differ by their virulence. We could detect byFlow cytometer that H3K76 could be mono, di and trimethylated, being the last one the most abundant mark inevery strain tested. This result is consistent with previous outcomes that showed that H3K76me3 is present inevery step of the cell cycle. Additionally, we could detect TcAUK1 in Dm28c, K98 and Tulahuen. This isimportant because the subcellular location and expression levels of this protein are finely regulated and it isevasive to Western blot detection.Furthermore, to check the potential connection between TcAUK1 and TcDOT1 activity, we are testing differentialH3K76 methylation in parasites that overexpress TcAUK1.Our findings surpass our study, since this approach will be useful for the whole community working intrypanosome epigenetics.Fil: López, María del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Lavignolle, María del Rosario. Centre D'immunologie Marseille-Luminy; FranciaFil: Vilchez Larrea, Salomé Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Ocampo, Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Argüello, Rafael. Centre D'immunologie Marseille-Luminy; FranciaFil: Alonso, Guillermo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaXXXVIII Annual Meeting of the Brazilian Society of Protozoology; XLIX Annual Meeting on Basic Research in Chagas’ DiseaseCaxambúBrasilBrazilian Society of ProtozoologyBrazilian Society of Protozoology2023info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/211751A novel approach to study epigenetic marks in Trypanosoma cruzi; XXXVIII Annual Meeting of the Brazilian Society of Protozoology; XLIX Annual Meeting on Basic Research in Chagas’ Disease; Caxambú; Brasil; 2023; 123-123CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://sbpz.org.br/wp-content/uploads/2023/09/livro-inteiro-04set.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 Técnicas2025-09-29T09:56:11Zoai:ri.conicet.gov.ar:11336/211751instacron: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 09:56:12.162CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A novel approach to study epigenetic marks in Trypanosoma cruzi
title A novel approach to study epigenetic marks in Trypanosoma cruzi
spellingShingle A novel approach to study epigenetic marks in Trypanosoma cruzi
López, María del Rosario
TRYPANOSOMATIDS
FLOW CITOMETRY
HISTONE METHYL TRANSFERASES
title_short A novel approach to study epigenetic marks in Trypanosoma cruzi
title_full A novel approach to study epigenetic marks in Trypanosoma cruzi
title_fullStr A novel approach to study epigenetic marks in Trypanosoma cruzi
title_full_unstemmed A novel approach to study epigenetic marks in Trypanosoma cruzi
title_sort A novel approach to study epigenetic marks in Trypanosoma cruzi
dc.creator.none.fl_str_mv López, María del Rosario
Lavignolle, María del Rosario
Vilchez Larrea, Salomé Catalina
Ocampo, Josefina
Argüello, Rafael
Alonso, Guillermo Daniel
author López, María del Rosario
author_facet López, María del Rosario
Lavignolle, María del Rosario
Vilchez Larrea, Salomé Catalina
Ocampo, Josefina
Argüello, Rafael
Alonso, Guillermo Daniel
author_role author
author2 Lavignolle, María del Rosario
Vilchez Larrea, Salomé Catalina
Ocampo, Josefina
Argüello, Rafael
Alonso, Guillermo Daniel
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv TRYPANOSOMATIDS
FLOW CITOMETRY
HISTONE METHYL TRANSFERASES
topic TRYPANOSOMATIDS
FLOW CITOMETRY
HISTONE METHYL TRANSFERASES
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv T. cruzi is the etiologic agent of Chagas disease and has a complex life cycle. Epimastigotes are the replicativeform in the vector, trypomastigotes are the infective stage, and amastigotes are a replicative intracellular stage.Proper cell cycle progression requires multiple factors.Epigenetics could be relevant for gene regulation. Particularly, lysine 76 of histone 3 (H3K76), could be mono,di or tri methylated by the methyltransferases TcDot1a and TcDot1b. It is not known how H3K76 methylationinfluences cell cycle progression, but deletion of TcDOT1b arrests the cell cycle in G2/M. Additionally,overexpression of Aurora kinase 1 (TcAUK1), affects the same stage.One of the main problems to study the epigenetic role of H3K76 methylation, or TcAUK1 in cell cycle progressionis that transgenic parasites for these proteins had been hard to obtain or maintain. Therefore, we used Flowcytometry that allowed us to identify the differential methylation of H3K76 and to make quantitative analysisusing low amounts of antibodies. Remarkably, we have been able to fine-tune the technique to this end. Wehave used three strain of T. cruzi, Dm28c; K98 and Tulahuen that differ by their virulence. We could detect byFlow cytometer that H3K76 could be mono, di and trimethylated, being the last one the most abundant mark inevery strain tested. This result is consistent with previous outcomes that showed that H3K76me3 is present inevery step of the cell cycle. Additionally, we could detect TcAUK1 in Dm28c, K98 and Tulahuen. This isimportant because the subcellular location and expression levels of this protein are finely regulated and it isevasive to Western blot detection.Furthermore, to check the potential connection between TcAUK1 and TcDOT1 activity, we are testing differentialH3K76 methylation in parasites that overexpress TcAUK1.Our findings surpass our study, since this approach will be useful for the whole community working intrypanosome epigenetics.
Fil: López, María del Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Lavignolle, María del Rosario. Centre D'immunologie Marseille-Luminy; Francia
Fil: Vilchez Larrea, Salomé Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Ocampo, Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Argüello, Rafael. Centre D'immunologie Marseille-Luminy; Francia
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
XXXVIII Annual Meeting of the Brazilian Society of Protozoology; XLIX Annual Meeting on Basic Research in Chagas’ Disease
Caxambú
Brasil
Brazilian Society of Protozoology
description T. cruzi is the etiologic agent of Chagas disease and has a complex life cycle. Epimastigotes are the replicativeform in the vector, trypomastigotes are the infective stage, and amastigotes are a replicative intracellular stage.Proper cell cycle progression requires multiple factors.Epigenetics could be relevant for gene regulation. Particularly, lysine 76 of histone 3 (H3K76), could be mono,di or tri methylated by the methyltransferases TcDot1a and TcDot1b. It is not known how H3K76 methylationinfluences cell cycle progression, but deletion of TcDOT1b arrests the cell cycle in G2/M. Additionally,overexpression of Aurora kinase 1 (TcAUK1), affects the same stage.One of the main problems to study the epigenetic role of H3K76 methylation, or TcAUK1 in cell cycle progressionis that transgenic parasites for these proteins had been hard to obtain or maintain. Therefore, we used Flowcytometry that allowed us to identify the differential methylation of H3K76 and to make quantitative analysisusing low amounts of antibodies. Remarkably, we have been able to fine-tune the technique to this end. Wehave used three strain of T. cruzi, Dm28c; K98 and Tulahuen that differ by their virulence. We could detect byFlow cytometer that H3K76 could be mono, di and trimethylated, being the last one the most abundant mark inevery strain tested. This result is consistent with previous outcomes that showed that H3K76me3 is present inevery step of the cell cycle. Additionally, we could detect TcAUK1 in Dm28c, K98 and Tulahuen. This isimportant because the subcellular location and expression levels of this protein are finely regulated and it isevasive to Western blot detection.Furthermore, to check the potential connection between TcAUK1 and TcDOT1 activity, we are testing differentialH3K76 methylation in parasites that overexpress TcAUK1.Our findings surpass our study, since this approach will be useful for the whole community working intrypanosome epigenetics.
publishDate 2023
dc.date.none.fl_str_mv 2023
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Book
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/11336/211751
A novel approach to study epigenetic marks in Trypanosoma cruzi; XXXVIII Annual Meeting of the Brazilian Society of Protozoology; XLIX Annual Meeting on Basic Research in Chagas’ Disease; Caxambú; Brasil; 2023; 123-123
CONICET Digital
CONICET
url http://hdl.handle.net/11336/211751
identifier_str_mv A novel approach to study epigenetic marks in Trypanosoma cruzi; XXXVIII Annual Meeting of the Brazilian Society of Protozoology; XLIX Annual Meeting on Basic Research in Chagas’ Disease; Caxambú; Brasil; 2023; 123-123
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://sbpz.org.br/wp-content/uploads/2023/09/livro-inteiro-04set.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
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Brazilian Society of Protozoology
publisher.none.fl_str_mv Brazilian Society of Protozoology
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 13.070432

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