Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi

Autores
De Lima, Loyze P.; Calderano, Simone G.; Da Silva, Mario Marcelo; de Araujo, Christiane B.; Vasconcelos, Elton J. R.; Iwai, Leo K.; Pereira, Claudio Alejandro; Fragoso, Stenio P.; Elias, Carolina M.
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
DNA polymerase theta (Polθ), a member of the DNA polymerase family A, exhibits a polymerase C-terminal domain, a central domain, and an N-terminal helicase domain. Polθ plays important roles in DNA repair via its polymerase domain, regulating genome integrity. In addition, in mammals, Polθ modulates origin firing timing and MCM helicase recruitment to chromatin. In contrast, as a model eukaryote, Trypanosoma cruzi exhibits two individual putative orthologs of Polθ in different genomic loci; one ortholog is homologous to the Polθ C-terminal polymerase domain, and the other is homologous to the Polθ helicase domain, called Polθ-polymerase and Polθ-helicase, respectively. A pull-down assay using the T. cruzi component of the prereplication complex Orc1/Cdc6 as bait captured Polθ-helicase from the nuclear extract. Orc1/Cdc6 and Polθ-helicase directly interacted, and Polθ-helicase presented DNA unwinding and ATPase activities. A T. cruzi strain overexpressing the Polθ-helicase domain exhibited a significantly decreased amount of DNA-bound MCM7 and impaired replication origin firing. Taken together, these data suggest that Polθ-helicase modulates DNA replication by directly interacting with Orc1/Cdc6, which reduces the binding of MCM7 to DNA and thereby impairs the firing of replication origins.
Fil: De Lima, Loyze P.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Calderano, Simone G.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Da Silva, Mario Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: de Araujo, Christiane B.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Vasconcelos, Elton J. R.. Western University Of Health Sciences.; Estados Unidos
Fil: Iwai, Leo K.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Pereira, Claudio Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina
Fil: Fragoso, Stenio P.. Instituto Carlos Chagas; Brasil
Fil: Elias, Carolina M.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Materia
TRYPANOSOMA CRUZI
DNA POLYMERASE THETA
HELICASE
CHAGAS DISEASE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/103148
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/103148
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruziDe Lima, Loyze P.Calderano, Simone G.Da Silva, Mario Marcelode Araujo, Christiane B.Vasconcelos, Elton J. R.Iwai, Leo K.Pereira, Claudio AlejandroFragoso, Stenio P.Elias, Carolina M.TRYPANOSOMA CRUZIDNA POLYMERASE THETAHELICASECHAGAS DISEASEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1DNA polymerase theta (Polθ), a member of the DNA polymerase family A, exhibits a polymerase C-terminal domain, a central domain, and an N-terminal helicase domain. Polθ plays important roles in DNA repair via its polymerase domain, regulating genome integrity. In addition, in mammals, Polθ modulates origin firing timing and MCM helicase recruitment to chromatin. In contrast, as a model eukaryote, Trypanosoma cruzi exhibits two individual putative orthologs of Polθ in different genomic loci; one ortholog is homologous to the Polθ C-terminal polymerase domain, and the other is homologous to the Polθ helicase domain, called Polθ-polymerase and Polθ-helicase, respectively. A pull-down assay using the T. cruzi component of the prereplication complex Orc1/Cdc6 as bait captured Polθ-helicase from the nuclear extract. Orc1/Cdc6 and Polθ-helicase directly interacted, and Polθ-helicase presented DNA unwinding and ATPase activities. A T. cruzi strain overexpressing the Polθ-helicase domain exhibited a significantly decreased amount of DNA-bound MCM7 and impaired replication origin firing. Taken together, these data suggest that Polθ-helicase modulates DNA replication by directly interacting with Orc1/Cdc6, which reduces the binding of MCM7 to DNA and thereby impairs the firing of replication origins.Fil: De Lima, Loyze P.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Calderano, Simone G.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Da Silva, Mario Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: de Araujo, Christiane B.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Vasconcelos, Elton J. R.. Western University Of Health Sciences.; Estados UnidosFil: Iwai, Leo K.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilFil: Pereira, Claudio Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Fragoso, Stenio P.. Instituto Carlos Chagas; BrasilFil: Elias, Carolina M.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; BrasilNature2019-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/103148De Lima, Loyze P.; Calderano, Simone G.; Da Silva, Mario Marcelo; de Araujo, Christiane B.; Vasconcelos, Elton J. R.; et al.; Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi; Nature; Scientific Reports; 9; 1; 2-2019; 1-162045-2322CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.nature.com/articles/s41598-019-39348-2info:eu-repo/semantics/altIdentifier/doi/10.1038/s41598-019-39348-2info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:04:20Zoai:ri.conicet.gov.ar:11336/103148instacron: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 10:04:20.44CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi
title Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi
spellingShingle Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi
De Lima, Loyze P.
TRYPANOSOMA CRUZI
DNA POLYMERASE THETA
HELICASE
CHAGAS DISEASE
title_short Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi
title_full Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi
title_fullStr Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi
title_full_unstemmed Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi
title_sort Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi
dc.creator.none.fl_str_mv De Lima, Loyze P.
Calderano, Simone G.
Da Silva, Mario Marcelo
de Araujo, Christiane B.
Vasconcelos, Elton J. R.
Iwai, Leo K.
Pereira, Claudio Alejandro
Fragoso, Stenio P.
Elias, Carolina M.
author De Lima, Loyze P.
author_facet De Lima, Loyze P.
Calderano, Simone G.
Da Silva, Mario Marcelo
de Araujo, Christiane B.
Vasconcelos, Elton J. R.
Iwai, Leo K.
Pereira, Claudio Alejandro
Fragoso, Stenio P.
Elias, Carolina M.
author_role author
author2 Calderano, Simone G.
Da Silva, Mario Marcelo
de Araujo, Christiane B.
Vasconcelos, Elton J. R.
Iwai, Leo K.
Pereira, Claudio Alejandro
Fragoso, Stenio P.
Elias, Carolina M.
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv TRYPANOSOMA CRUZI
DNA POLYMERASE THETA
HELICASE
CHAGAS DISEASE
topic TRYPANOSOMA CRUZI
DNA POLYMERASE THETA
HELICASE
CHAGAS DISEASE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv DNA polymerase theta (Polθ), a member of the DNA polymerase family A, exhibits a polymerase C-terminal domain, a central domain, and an N-terminal helicase domain. Polθ plays important roles in DNA repair via its polymerase domain, regulating genome integrity. In addition, in mammals, Polθ modulates origin firing timing and MCM helicase recruitment to chromatin. In contrast, as a model eukaryote, Trypanosoma cruzi exhibits two individual putative orthologs of Polθ in different genomic loci; one ortholog is homologous to the Polθ C-terminal polymerase domain, and the other is homologous to the Polθ helicase domain, called Polθ-polymerase and Polθ-helicase, respectively. A pull-down assay using the T. cruzi component of the prereplication complex Orc1/Cdc6 as bait captured Polθ-helicase from the nuclear extract. Orc1/Cdc6 and Polθ-helicase directly interacted, and Polθ-helicase presented DNA unwinding and ATPase activities. A T. cruzi strain overexpressing the Polθ-helicase domain exhibited a significantly decreased amount of DNA-bound MCM7 and impaired replication origin firing. Taken together, these data suggest that Polθ-helicase modulates DNA replication by directly interacting with Orc1/Cdc6, which reduces the binding of MCM7 to DNA and thereby impairs the firing of replication origins.
Fil: De Lima, Loyze P.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Calderano, Simone G.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Da Silva, Mario Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: de Araujo, Christiane B.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Vasconcelos, Elton J. R.. Western University Of Health Sciences.; Estados Unidos
Fil: Iwai, Leo K.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
Fil: Pereira, Claudio Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina
Fil: Fragoso, Stenio P.. Instituto Carlos Chagas; Brasil
Fil: Elias, Carolina M.. Governo do Estado de Sao Paulo. Secretaria da Saude. Instituto Butantan; Brasil
description DNA polymerase theta (Polθ), a member of the DNA polymerase family A, exhibits a polymerase C-terminal domain, a central domain, and an N-terminal helicase domain. Polθ plays important roles in DNA repair via its polymerase domain, regulating genome integrity. In addition, in mammals, Polθ modulates origin firing timing and MCM helicase recruitment to chromatin. In contrast, as a model eukaryote, Trypanosoma cruzi exhibits two individual putative orthologs of Polθ in different genomic loci; one ortholog is homologous to the Polθ C-terminal polymerase domain, and the other is homologous to the Polθ helicase domain, called Polθ-polymerase and Polθ-helicase, respectively. A pull-down assay using the T. cruzi component of the prereplication complex Orc1/Cdc6 as bait captured Polθ-helicase from the nuclear extract. Orc1/Cdc6 and Polθ-helicase directly interacted, and Polθ-helicase presented DNA unwinding and ATPase activities. A T. cruzi strain overexpressing the Polθ-helicase domain exhibited a significantly decreased amount of DNA-bound MCM7 and impaired replication origin firing. Taken together, these data suggest that Polθ-helicase modulates DNA replication by directly interacting with Orc1/Cdc6, which reduces the binding of MCM7 to DNA and thereby impairs the firing of replication origins.
publishDate 2019
dc.date.none.fl_str_mv 2019-02
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/11336/103148
De Lima, Loyze P.; Calderano, Simone G.; Da Silva, Mario Marcelo; de Araujo, Christiane B.; Vasconcelos, Elton J. R.; et al.; Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi; Nature; Scientific Reports; 9; 1; 2-2019; 1-16
2045-2322
CONICET Digital
CONICET
url http://hdl.handle.net/11336/103148
identifier_str_mv De Lima, Loyze P.; Calderano, Simone G.; Da Silva, Mario Marcelo; de Araujo, Christiane B.; Vasconcelos, Elton J. R.; et al.; Ortholog of the polymerase theta helicase domain modulates DNA replication in Trypanosoma cruzi; Nature; Scientific Reports; 9; 1; 2-2019; 1-16
2045-2322
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.nature.com/articles/s41598-019-39348-2
info:eu-repo/semantics/altIdentifier/doi/10.1038/s41598-019-39348-2
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Nature
publisher.none.fl_str_mv Nature
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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