Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi

Autores
Rodríguez Durán, Jessica Jenireth; Gomez, Karina Andrea; Potenza, Mariana
Año de publicación
2019
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Calcium homeostasis is implicated in essential processes of T. cruzilife cycle. Although several calcium binding proteins have beenidentified in T. cruzi genome, their role in Ca2+ signaling remain stillunknown. TcCAL1 is a hypothetical protein identified throughproteome analysis of T. cruzi, which has two EF-Hand domainsinvolved in Ca2+ binding. The hypothesis to be tested in this studywas that TcCAL1 overexpression could trigger alterations in theintracellular calcium homeostasis leading an effect on epimastigoteproliferation. For this purpose, the gene encoding tccal1 wasamplified by PCR and cloned fussed to a 6x histidine tag into thepTREX expression vector. The recombinant plasmidpTREX/tccal1x6His was used to transfect epimastigotes from T.cruzi Y or CL Brener strains. Selection of transgenic cultures wascarried out growing parasites in presence of increasingconcentrations of Geneticin. Overexpression of TcCAL1x6His fusionprotein was confirmed through western blot and affinitychromatography. Growth curves of recombinant cultures from Y orCL Brener strains were performed in triplicate, counting theparasites in a Neubauer chamber. Parasite cultures carrying theempty vector pTREX were used as controls. Data obtained wereprocessed using GraphPad Prism 6.0 Software and subjected tostatistical analysis by two-way ANOVA test. The preliminary resultsshowed that the overexpression of TcCAL1x6His affects the growthcurve of CL Brenner strain at the stationary phase but theproliferation Y strain is not affected. Additional experiments arebeing carrying out to validate these results and to study the effectof TcCAL1 overexpression on T. cruzi differentiation. Future assaysincluding parasites carrying the tccal1 gene silenced will contributeto validate our hypothesis and to reveal the function of TcCAL1 incalcium homeostasis of T. cruzi.
Fil: Rodríguez Durán, Jessica Jenireth. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Gomez, Karina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Potenza, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Mar del Plata
Argentina
Sociedad Argentina de Investigación Clínica
Asociación Argentina de Farmacología Experimenta
Sociedad Argentina de Biología
Sociedad Argentina de Protozoología
Asociación Argentina de Nanomedicinas
Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Materia
Trypanosoma cruzi
Calcium binding
Parasitology
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:11336/201594

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network_acronym_str CONICETDig
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network_name_str CONICET Digital (CONICET)
spelling Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruziRodríguez Durán, Jessica JenirethGomez, Karina AndreaPotenza, MarianaTrypanosoma cruziCalcium bindingParasitologyhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Calcium homeostasis is implicated in essential processes of T. cruzilife cycle. Although several calcium binding proteins have beenidentified in T. cruzi genome, their role in Ca2+ signaling remain stillunknown. TcCAL1 is a hypothetical protein identified throughproteome analysis of T. cruzi, which has two EF-Hand domainsinvolved in Ca2+ binding. The hypothesis to be tested in this studywas that TcCAL1 overexpression could trigger alterations in theintracellular calcium homeostasis leading an effect on epimastigoteproliferation. For this purpose, the gene encoding tccal1 wasamplified by PCR and cloned fussed to a 6x histidine tag into thepTREX expression vector. The recombinant plasmidpTREX/tccal1x6His was used to transfect epimastigotes from T.cruzi Y or CL Brener strains. Selection of transgenic cultures wascarried out growing parasites in presence of increasingconcentrations of Geneticin. Overexpression of TcCAL1x6His fusionprotein was confirmed through western blot and affinitychromatography. Growth curves of recombinant cultures from Y orCL Brener strains were performed in triplicate, counting theparasites in a Neubauer chamber. Parasite cultures carrying theempty vector pTREX were used as controls. Data obtained wereprocessed using GraphPad Prism 6.0 Software and subjected tostatistical analysis by two-way ANOVA test. The preliminary resultsshowed that the overexpression of TcCAL1x6His affects the growthcurve of CL Brenner strain at the stationary phase but theproliferation Y strain is not affected. Additional experiments arebeing carrying out to validate these results and to study the effectof TcCAL1 overexpression on T. cruzi differentiation. Future assaysincluding parasites carrying the tccal1 gene silenced will contributeto validate our hypothesis and to reveal the function of TcCAL1 incalcium homeostasis of T. cruzi.Fil: Rodríguez Durán, Jessica Jenireth. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Gomez, Karina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Potenza, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaLXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de LaboratorioMar del PlataArgentinaSociedad Argentina de Investigación ClínicaAsociación Argentina de Farmacología ExperimentaSociedad Argentina de BiologíaSociedad Argentina de ProtozoologíaAsociación Argentina de NanomedicinasAsociación Argentina de Ciencia y Tecnología de Animales de LaboratorioFundación Revista MedicinaCostas, Monica AlejandraMarino, Gabriela InésAzurmendi, Pablo Javier2019info: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/11336/201594Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio; Mar del Plata; Argentina; 2019; 121-1210025-76801669-9106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://medicinabuenosaires.com/revistas/vol79-19/s4/vol79_s4.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 Técnicas2025-10-15T15:03:46Zoai:ri.conicet.gov.ar:11336/201594instacron: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 15:03:46.562CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi
title Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi
spellingShingle Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi
Rodríguez Durán, Jessica Jenireth
Trypanosoma cruzi
Calcium binding
Parasitology
title_short Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi
title_full Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi
title_fullStr Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi
title_full_unstemmed Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi
title_sort Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi
dc.creator.none.fl_str_mv Rodríguez Durán, Jessica Jenireth
Gomez, Karina Andrea
Potenza, Mariana
author Rodríguez Durán, Jessica Jenireth
author_facet Rodríguez Durán, Jessica Jenireth
Gomez, Karina Andrea
Potenza, Mariana
author_role author
author2 Gomez, Karina Andrea
Potenza, Mariana
author2_role author
author
dc.contributor.none.fl_str_mv Costas, Monica Alejandra
Marino, Gabriela Inés
Azurmendi, Pablo Javier
dc.subject.none.fl_str_mv Trypanosoma cruzi
Calcium binding
Parasitology
topic Trypanosoma cruzi
Calcium binding
Parasitology
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Calcium homeostasis is implicated in essential processes of T. cruzilife cycle. Although several calcium binding proteins have beenidentified in T. cruzi genome, their role in Ca2+ signaling remain stillunknown. TcCAL1 is a hypothetical protein identified throughproteome analysis of T. cruzi, which has two EF-Hand domainsinvolved in Ca2+ binding. The hypothesis to be tested in this studywas that TcCAL1 overexpression could trigger alterations in theintracellular calcium homeostasis leading an effect on epimastigoteproliferation. For this purpose, the gene encoding tccal1 wasamplified by PCR and cloned fussed to a 6x histidine tag into thepTREX expression vector. The recombinant plasmidpTREX/tccal1x6His was used to transfect epimastigotes from T.cruzi Y or CL Brener strains. Selection of transgenic cultures wascarried out growing parasites in presence of increasingconcentrations of Geneticin. Overexpression of TcCAL1x6His fusionprotein was confirmed through western blot and affinitychromatography. Growth curves of recombinant cultures from Y orCL Brener strains were performed in triplicate, counting theparasites in a Neubauer chamber. Parasite cultures carrying theempty vector pTREX were used as controls. Data obtained wereprocessed using GraphPad Prism 6.0 Software and subjected tostatistical analysis by two-way ANOVA test. The preliminary resultsshowed that the overexpression of TcCAL1x6His affects the growthcurve of CL Brenner strain at the stationary phase but theproliferation Y strain is not affected. Additional experiments arebeing carrying out to validate these results and to study the effectof TcCAL1 overexpression on T. cruzi differentiation. Future assaysincluding parasites carrying the tccal1 gene silenced will contributeto validate our hypothesis and to reveal the function of TcCAL1 incalcium homeostasis of T. cruzi.
Fil: Rodríguez Durán, Jessica Jenireth. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Gomez, Karina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Potenza, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
Mar del Plata
Argentina
Sociedad Argentina de Investigación Clínica
Asociación Argentina de Farmacología Experimenta
Sociedad Argentina de Biología
Sociedad Argentina de Protozoología
Asociación Argentina de Nanomedicinas
Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio
description Calcium homeostasis is implicated in essential processes of T. cruzilife cycle. Although several calcium binding proteins have beenidentified in T. cruzi genome, their role in Ca2+ signaling remain stillunknown. TcCAL1 is a hypothetical protein identified throughproteome analysis of T. cruzi, which has two EF-Hand domainsinvolved in Ca2+ binding. The hypothesis to be tested in this studywas that TcCAL1 overexpression could trigger alterations in theintracellular calcium homeostasis leading an effect on epimastigoteproliferation. For this purpose, the gene encoding tccal1 wasamplified by PCR and cloned fussed to a 6x histidine tag into thepTREX expression vector. The recombinant plasmidpTREX/tccal1x6His was used to transfect epimastigotes from T.cruzi Y or CL Brener strains. Selection of transgenic cultures wascarried out growing parasites in presence of increasingconcentrations of Geneticin. Overexpression of TcCAL1x6His fusionprotein was confirmed through western blot and affinitychromatography. Growth curves of recombinant cultures from Y orCL Brener strains were performed in triplicate, counting theparasites in a Neubauer chamber. Parasite cultures carrying theempty vector pTREX were used as controls. Data obtained wereprocessed using GraphPad Prism 6.0 Software and subjected tostatistical analysis by two-way ANOVA test. The preliminary resultsshowed that the overexpression of TcCAL1x6His affects the growthcurve of CL Brenner strain at the stationary phase but theproliferation Y strain is not affected. Additional experiments arebeing carrying out to validate these results and to study the effectof TcCAL1 overexpression on T. cruzi differentiation. Future assaysincluding parasites carrying the tccal1 gene silenced will contributeto validate our hypothesis and to reveal the function of TcCAL1 incalcium homeostasis of T. 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
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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/11336/201594
Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio; Mar del Plata; Argentina; 2019; 121-121
0025-7680
1669-9106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/201594
identifier_str_mv Effect of the calcium binding protein TcCAL1 on the proliferation of Trypanosoma cruzi; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimenta; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de la Asociación Argentina de Nanomedicinas; VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de Laboratorio; Mar del Plata; Argentina; 2019; 121-121
0025-7680
1669-9106
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://medicinabuenosaires.com/revistas/vol79-19/s4/vol79_s4.pdf
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dc.coverage.none.fl_str_mv Internacional
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publisher.none.fl_str_mv Fundación Revista Medicina
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