Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle

Autores
Balestrasse, Malena; Massimino Stepñicka, Milena; Beati, Maria Paula; Alonso, Guillermo Daniel; Ocampo, Josefina
Año de publicación
2020
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Trypanosoma cruzi, the etiologic agent of Chagas Disease, affects a large number of the population in Latin America. It has a complex life cycle alternating between a mammalian host and the vector insect, Triatoma infestans. This cycle consists of three well-defined stages: amastigotes, epimastigotes and trypomastigotes. As the parasite faces different environments, it requires changes in gene expression in order to survive. Hence, gene expression regulation might be a key aspect to understand adaptation. Despite Trypanosomes gene expression is mainly regulated post transcriptionally, there are evidences that chromatin state influence it. In T. cruzi, there are two Dot1 methyltransferases homologues, called Dot1a and Dot1b, involved in the sequential mono, di and tri methylation of lysine 76 of histone H3 (H3K76me). Additionally, recent studies have shown that Dot1a deletion is not viable while Dot1b null mutant has aberrant morphology and decreased growth rate.In this project, we investigated the relevance of TcDot1a and TcDot1b during cell cycle and the metacyclogenesis process. Therefore, to analyze the isoforms subcelular location and their effects on cell cycle progression and differentiation, we have cloned the TcDot1 isoforms in a pRibotex vector with an N-terminal HA-tag and transfected epimastigotes of CL Brener strain. Nevertheless, the overexpression was toxic for the cell. Therefore, we decided to switch to an inducible vector.In order to elucidate the epigenetic implications of H3K76 differential methylation in the different stages of the life cycle of T. cruzi, we propose to map H3K76 mono, di and tri-methylation genome wide by MNase-ChIP-seq technique followed by paired-end sequencing.Overall, our data will be useful to further understand the role of the Dot1 isoforms and the epigenetic mechanism mediated by H3K76 in this unique parasite.
Fil: Balestrasse, Malena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Massimino Stepñicka, Milena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Beati, Maria Paula. 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
Fil: Ocampo, Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
XXXII Reunión Anual de la Sociedad Argentina de Protozoología
Ciudad Autónoma de Buenos Aires
Argentina
Sociedad Argentina de Protozoología
Materia
Chromatin
TcDot1
Trupanosoma cruzi
H3K76
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:11336/201803
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/201803
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycleBalestrasse, MalenaMassimino Stepñicka, MilenaBeati, Maria PaulaAlonso, Guillermo DanielOcampo, JosefinaChromatinTcDot1Trupanosoma cruziH3K76https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Trypanosoma cruzi, the etiologic agent of Chagas Disease, affects a large number of the population in Latin America. It has a complex life cycle alternating between a mammalian host and the vector insect, Triatoma infestans. This cycle consists of three well-defined stages: amastigotes, epimastigotes and trypomastigotes. As the parasite faces different environments, it requires changes in gene expression in order to survive. Hence, gene expression regulation might be a key aspect to understand adaptation. Despite Trypanosomes gene expression is mainly regulated post transcriptionally, there are evidences that chromatin state influence it. In T. cruzi, there are two Dot1 methyltransferases homologues, called Dot1a and Dot1b, involved in the sequential mono, di and tri methylation of lysine 76 of histone H3 (H3K76me). Additionally, recent studies have shown that Dot1a deletion is not viable while Dot1b null mutant has aberrant morphology and decreased growth rate.In this project, we investigated the relevance of TcDot1a and TcDot1b during cell cycle and the metacyclogenesis process. Therefore, to analyze the isoforms subcelular location and their effects on cell cycle progression and differentiation, we have cloned the TcDot1 isoforms in a pRibotex vector with an N-terminal HA-tag and transfected epimastigotes of CL Brener strain. Nevertheless, the overexpression was toxic for the cell. Therefore, we decided to switch to an inducible vector.In order to elucidate the epigenetic implications of H3K76 differential methylation in the different stages of the life cycle of T. cruzi, we propose to map H3K76 mono, di and tri-methylation genome wide by MNase-ChIP-seq technique followed by paired-end sequencing.Overall, our data will be useful to further understand the role of the Dot1 isoforms and the epigenetic mechanism mediated by H3K76 in this unique parasite.Fil: Balestrasse, Malena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Massimino Stepñicka, Milena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Beati, Maria Paula. 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"; ArgentinaFil: Ocampo, Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaXXXII Reunión Anual de la Sociedad Argentina de ProtozoologíaCiudad Autónoma de Buenos AiresArgentinaSociedad Argentina de ProtozoologíaSociedad Argentina de ProtozoologíaRiarte, Adelina RosaLonghi, Silvia AndreaFrank, Fernanda María2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/201803Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle; XXXII Reunión Anual de la Sociedad Argentina de Protozoología; Ciudad Autónoma de Buenos Aires; Argentina; 2020; 55-55CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://protozoologia.org.ar/wp-content/uploads/Libro-de-Resumenes-SAP-2020.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:42:43Zoai:ri.conicet.gov.ar:11336/201803instacron: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 15:42:43.713CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle
title Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle
spellingShingle Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle
Balestrasse, Malena
Chromatin
TcDot1
Trupanosoma cruzi
H3K76
title_short Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle
title_full Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle
title_fullStr Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle
title_full_unstemmed Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle
title_sort Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle
dc.creator.none.fl_str_mv Balestrasse, Malena
Massimino Stepñicka, Milena
Beati, Maria Paula
Alonso, Guillermo Daniel
Ocampo, Josefina
author Balestrasse, Malena
author_facet Balestrasse, Malena
Massimino Stepñicka, Milena
Beati, Maria Paula
Alonso, Guillermo Daniel
Ocampo, Josefina
author_role author
author2 Massimino Stepñicka, Milena
Beati, Maria Paula
Alonso, Guillermo Daniel
Ocampo, Josefina
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Riarte, Adelina Rosa
Longhi, Silvia Andrea
Frank, Fernanda María
dc.subject.none.fl_str_mv Chromatin
TcDot1
Trupanosoma cruzi
H3K76
topic Chromatin
TcDot1
Trupanosoma cruzi
H3K76
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Trypanosoma cruzi, the etiologic agent of Chagas Disease, affects a large number of the population in Latin America. It has a complex life cycle alternating between a mammalian host and the vector insect, Triatoma infestans. This cycle consists of three well-defined stages: amastigotes, epimastigotes and trypomastigotes. As the parasite faces different environments, it requires changes in gene expression in order to survive. Hence, gene expression regulation might be a key aspect to understand adaptation. Despite Trypanosomes gene expression is mainly regulated post transcriptionally, there are evidences that chromatin state influence it. In T. cruzi, there are two Dot1 methyltransferases homologues, called Dot1a and Dot1b, involved in the sequential mono, di and tri methylation of lysine 76 of histone H3 (H3K76me). Additionally, recent studies have shown that Dot1a deletion is not viable while Dot1b null mutant has aberrant morphology and decreased growth rate.In this project, we investigated the relevance of TcDot1a and TcDot1b during cell cycle and the metacyclogenesis process. Therefore, to analyze the isoforms subcelular location and their effects on cell cycle progression and differentiation, we have cloned the TcDot1 isoforms in a pRibotex vector with an N-terminal HA-tag and transfected epimastigotes of CL Brener strain. Nevertheless, the overexpression was toxic for the cell. Therefore, we decided to switch to an inducible vector.In order to elucidate the epigenetic implications of H3K76 differential methylation in the different stages of the life cycle of T. cruzi, we propose to map H3K76 mono, di and tri-methylation genome wide by MNase-ChIP-seq technique followed by paired-end sequencing.Overall, our data will be useful to further understand the role of the Dot1 isoforms and the epigenetic mechanism mediated by H3K76 in this unique parasite.
Fil: Balestrasse, Malena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Massimino Stepñicka, Milena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Beati, Maria Paula. 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
Fil: Ocampo, Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
XXXII Reunión Anual de la Sociedad Argentina de Protozoología
Ciudad Autónoma de Buenos Aires
Argentina
Sociedad Argentina de Protozoología
description Trypanosoma cruzi, the etiologic agent of Chagas Disease, affects a large number of the population in Latin America. It has a complex life cycle alternating between a mammalian host and the vector insect, Triatoma infestans. This cycle consists of three well-defined stages: amastigotes, epimastigotes and trypomastigotes. As the parasite faces different environments, it requires changes in gene expression in order to survive. Hence, gene expression regulation might be a key aspect to understand adaptation. Despite Trypanosomes gene expression is mainly regulated post transcriptionally, there are evidences that chromatin state influence it. In T. cruzi, there are two Dot1 methyltransferases homologues, called Dot1a and Dot1b, involved in the sequential mono, di and tri methylation of lysine 76 of histone H3 (H3K76me). Additionally, recent studies have shown that Dot1a deletion is not viable while Dot1b null mutant has aberrant morphology and decreased growth rate.In this project, we investigated the relevance of TcDot1a and TcDot1b during cell cycle and the metacyclogenesis process. Therefore, to analyze the isoforms subcelular location and their effects on cell cycle progression and differentiation, we have cloned the TcDot1 isoforms in a pRibotex vector with an N-terminal HA-tag and transfected epimastigotes of CL Brener strain. Nevertheless, the overexpression was toxic for the cell. Therefore, we decided to switch to an inducible vector.In order to elucidate the epigenetic implications of H3K76 differential methylation in the different stages of the life cycle of T. cruzi, we propose to map H3K76 mono, di and tri-methylation genome wide by MNase-ChIP-seq technique followed by paired-end sequencing.Overall, our data will be useful to further understand the role of the Dot1 isoforms and the epigenetic mechanism mediated by H3K76 in this unique parasite.
publishDate 2020
dc.date.none.fl_str_mv 2020
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/11336/201803
Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle; XXXII Reunión Anual de la Sociedad Argentina de Protozoología; Ciudad Autónoma de Buenos Aires; Argentina; 2020; 55-55
CONICET Digital
CONICET
url http://hdl.handle.net/11336/201803
identifier_str_mv Role of the TcDOT1a and TcDOT1b isoforms in H3K76 differential methylation and their impact in Trypanosoma cruzi life cycle; XXXII Reunión Anual de la Sociedad Argentina de Protozoología; Ciudad Autónoma de Buenos Aires; Argentina; 2020; 55-55
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://protozoologia.org.ar/wp-content/uploads/Libro-de-Resumenes-SAP-2020.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
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Sociedad Argentina de Protozoología
publisher.none.fl_str_mv Sociedad Argentina de Protozoología
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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