Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding Protein

Detalhes bibliográficos
Autor(a) principal: Rodrigues, Ana P. C. [UNESP]
Data de Publicação: 2021
Outros Autores: Oliveira, Marcos T. [UNESP]
Tipo de documento: Capítulo de livro
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1007/978-1-0716-1290-3_20
http://hdl.handle.net/11449/228928
Resumo: Defects in mitochondrial DNA (mtDNA) maintenance may lead to disturbances in mitochondrial homeostasis and energy production in eukaryotic cells, causing diseases. During mtDNA replication, the mitochondrial single-stranded DNA-binding protein (mtSSB) stabilizes and protects the exposed single-stranded mtDNA from nucleolysis; perhaps more importantly, it appears to coordinate the actions of both the replicative mtDNA helicase Twinkle and DNA polymerase gamma at the replication fork. Here, we describe a helicase stimulation protocol to test in vitro the functional interaction between mtSSB and variant forms of Twinkle. We show for the first time that the C-terminal tail of Twinkle is important for such an interaction, and that it negatively regulates helicase unwinding activity in a salt-dependent manner.
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spelling Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding ProteindsDNA unwinding assayMitochondrial DNA replicationmtSSBP66TwinkleDefects in mitochondrial DNA (mtDNA) maintenance may lead to disturbances in mitochondrial homeostasis and energy production in eukaryotic cells, causing diseases. During mtDNA replication, the mitochondrial single-stranded DNA-binding protein (mtSSB) stabilizes and protects the exposed single-stranded mtDNA from nucleolysis; perhaps more importantly, it appears to coordinate the actions of both the replicative mtDNA helicase Twinkle and DNA polymerase gamma at the replication fork. Here, we describe a helicase stimulation protocol to test in vitro the functional interaction between mtSSB and variant forms of Twinkle. We show for the first time that the C-terminal tail of Twinkle is important for such an interaction, and that it negatively regulates helicase unwinding activity in a salt-dependent manner.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Departamento de Tecnologia Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista “Júlio de Mesquita Filho”Departamento de Tecnologia Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista “Júlio de Mesquita Filho”CNPq: 140788/2018-2FAPESP: 2017/ 04372-0CNPq: 306974/2017-7CNPq: 424562/2018-9Universidade Estadual Paulista (UNESP)Rodrigues, Ana P. C. [UNESP]Oliveira, Marcos T. [UNESP]2022-04-29T08:29:27Z2022-04-29T08:29:27Z2021-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/bookPart313-322http://dx.doi.org/10.1007/978-1-0716-1290-3_20Methods in Molecular Biology, v. 2281, p. 313-322.1940-60291064-3745http://hdl.handle.net/11449/22892810.1007/978-1-0716-1290-3_202-s2.0-85104297700Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMethods in Molecular Biologyinfo:eu-repo/semantics/openAccess2022-04-29T08:29:28Zoai:repositorio.unesp.br:11449/228928Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462022-04-29T08:29:28Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding Protein
title Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding Protein
spellingShingle Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding Protein
Rodrigues, Ana P. C. [UNESP]
dsDNA unwinding assay
Mitochondrial DNA replication
mtSSB
P66
Twinkle
title_short Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding Protein
title_full Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding Protein
title_fullStr Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding Protein
title_full_unstemmed Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding Protein
title_sort Stimulation of Variant Forms of the Mitochondrial DNA Helicase Twinkle by the Mitochondrial Single-Stranded DNA-Binding Protein
author Rodrigues, Ana P. C. [UNESP]
author_facet Rodrigues, Ana P. C. [UNESP]
Oliveira, Marcos T. [UNESP]
author_role author
author2 Oliveira, Marcos T. [UNESP]
author2_role author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (UNESP)
dc.contributor.author.fl_str_mv Rodrigues, Ana P. C. [UNESP]
Oliveira, Marcos T. [UNESP]
dc.subject.por.fl_str_mv dsDNA unwinding assay
Mitochondrial DNA replication
mtSSB
P66
Twinkle
topic dsDNA unwinding assay
Mitochondrial DNA replication
mtSSB
P66
Twinkle
description Defects in mitochondrial DNA (mtDNA) maintenance may lead to disturbances in mitochondrial homeostasis and energy production in eukaryotic cells, causing diseases. During mtDNA replication, the mitochondrial single-stranded DNA-binding protein (mtSSB) stabilizes and protects the exposed single-stranded mtDNA from nucleolysis; perhaps more importantly, it appears to coordinate the actions of both the replicative mtDNA helicase Twinkle and DNA polymerase gamma at the replication fork. Here, we describe a helicase stimulation protocol to test in vitro the functional interaction between mtSSB and variant forms of Twinkle. We show for the first time that the C-terminal tail of Twinkle is important for such an interaction, and that it negatively regulates helicase unwinding activity in a salt-dependent manner.
publishDate 2021
dc.date.none.fl_str_mv 2021-01-01
2022-04-29T08:29:27Z
2022-04-29T08:29:27Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/bookPart
format bookPart
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1007/978-1-0716-1290-3_20
Methods in Molecular Biology, v. 2281, p. 313-322.
1940-6029
1064-3745
http://hdl.handle.net/11449/228928
10.1007/978-1-0716-1290-3_20
2-s2.0-85104297700
url http://dx.doi.org/10.1007/978-1-0716-1290-3_20
http://hdl.handle.net/11449/228928
identifier_str_mv Methods in Molecular Biology, v. 2281, p. 313-322.
1940-6029
1064-3745
10.1007/978-1-0716-1290-3_20
2-s2.0-85104297700
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Methods in Molecular Biology
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 313-322
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
_version_ 1799965716820000768

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