AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements

Autores
Fernandez, Maricruz; Shkumatov, Alexander V.; Liu, Yun; Stulemeijer, Claire; Derclaye, Sylvie; Efremov, Rouslan G.; Hallet, Bernard; Alsteens, David
Año de publicación
2023
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Transposon Tn4430 belongs to a widespread family of bacterial transposons, the Tn3 family, whichplays a prevalent role in the dissemination of antibiotic resistance among pathogens. Despite recentdata on the structural architecture of the transposition complex, the molecular mechanisms underlyingthe replicative transposition of these elements arestill poorly understood. Here, we use force-distancecurve-based atomic force microscopy to probe thebinding of the TnpA transposase of Tn4430 to DNAmolecules containing one or two transposon endsand to extract the thermodynamic and kinetic parameters of transposition complex assembly. Comparing wild-type TnpA with previously isolated deregulated TnpA mutants supports a stepwise pathwayfor transposition complex formation and activationduring which TnpA first binds as a dimer to a single transposon end and then undergoes a structuraltransition that enables it to bind the second end cooperatively and to become activated for transpositioncatalysis, the latter step occurring at a much fasterrate for the TnpA mutants. Our study thus providesan unprecedented approach to probe the dynamic ofa complex DNA processing machinery at the singleparticle level.
Fil: Fernandez, Maricruz. Université Catholique de Louvain; Bélgica. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigaciones en Tecnologías Energéticas y Materiales Avanzados. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Tecnologías Energéticas y Materiales Avanzados; Argentina
Fil: Shkumatov, Alexander V.. Faculty Of Sciences And Bioengineering Sciences ; Vrije Universiteit Brussel;
Fil: Liu, Yun. Faculty Of Sciences And Bioengineering Sciences ; Vrije Universiteit Brussel;
Fil: Stulemeijer, Claire. Université Catholique de Louvain; Bélgica
Fil: Derclaye, Sylvie. Université Catholique de Louvain; Bélgica
Fil: Efremov, Rouslan G.. Faculty Of Sciences And Bioengineering Sciences ; Vrije Universiteit Brussel;
Fil: Hallet, Bernard. Université Catholique de Louvain; Bélgica
Fil: Alsteens, David. Université Catholique de Louvain; Bélgica
Materia
TRANSPOSITION
PROTEIN-DNA INTERACTION
DYNAMIC FORCE SPECTROSCOPY
AFM
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:11336/229930
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/229930
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elementsFernandez, MaricruzShkumatov, Alexander V.Liu, YunStulemeijer, ClaireDerclaye, SylvieEfremov, Rouslan G.Hallet, BernardAlsteens, DavidTRANSPOSITIONPROTEIN-DNA INTERACTIONDYNAMIC FORCE SPECTROSCOPYAFMhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Transposon Tn4430 belongs to a widespread family of bacterial transposons, the Tn3 family, whichplays a prevalent role in the dissemination of antibiotic resistance among pathogens. Despite recentdata on the structural architecture of the transposition complex, the molecular mechanisms underlyingthe replicative transposition of these elements arestill poorly understood. Here, we use force-distancecurve-based atomic force microscopy to probe thebinding of the TnpA transposase of Tn4430 to DNAmolecules containing one or two transposon endsand to extract the thermodynamic and kinetic parameters of transposition complex assembly. Comparing wild-type TnpA with previously isolated deregulated TnpA mutants supports a stepwise pathwayfor transposition complex formation and activationduring which TnpA first binds as a dimer to a single transposon end and then undergoes a structuraltransition that enables it to bind the second end cooperatively and to become activated for transpositioncatalysis, the latter step occurring at a much fasterrate for the TnpA mutants. Our study thus providesan unprecedented approach to probe the dynamic ofa complex DNA processing machinery at the singleparticle level.Fil: Fernandez, Maricruz. Université Catholique de Louvain; Bélgica. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigaciones en Tecnologías Energéticas y Materiales Avanzados. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Tecnologías Energéticas y Materiales Avanzados; ArgentinaFil: Shkumatov, Alexander V.. Faculty Of Sciences And Bioengineering Sciences ; Vrije Universiteit Brussel;Fil: Liu, Yun. Faculty Of Sciences And Bioengineering Sciences ; Vrije Universiteit Brussel;Fil: Stulemeijer, Claire. Université Catholique de Louvain; BélgicaFil: Derclaye, Sylvie. Université Catholique de Louvain; BélgicaFil: Efremov, Rouslan G.. Faculty Of Sciences And Bioengineering Sciences ; Vrije Universiteit Brussel;Fil: Hallet, Bernard. Université Catholique de Louvain; BélgicaFil: Alsteens, David. Université Catholique de Louvain; BélgicaOxford University Press2023-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/229930Fernandez, Maricruz; Shkumatov, Alexander V.; Liu, Yun; Stulemeijer, Claire; Derclaye, Sylvie; et al.; AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements; Oxford University Press; Nucleic Acids Research; 51; 10; 4-2023; 4929-49410305-10481362-4962CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://academic.oup.com/nar/advance-article/doi/10.1093/nar/gkad241/7110757info:eu-repo/semantics/altIdentifier/doi/10.1093/nar/gkad241info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:24:17Zoai:ri.conicet.gov.ar:11336/229930instacron: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 10:24:17.436CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements
title AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements
spellingShingle AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements
Fernandez, Maricruz
TRANSPOSITION
PROTEIN-DNA INTERACTION
DYNAMIC FORCE SPECTROSCOPY
AFM
title_short AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements
title_full AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements
title_fullStr AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements
title_full_unstemmed AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements
title_sort AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements
dc.creator.none.fl_str_mv Fernandez, Maricruz
Shkumatov, Alexander V.
Liu, Yun
Stulemeijer, Claire
Derclaye, Sylvie
Efremov, Rouslan G.
Hallet, Bernard
Alsteens, David
author Fernandez, Maricruz
author_facet Fernandez, Maricruz
Shkumatov, Alexander V.
Liu, Yun
Stulemeijer, Claire
Derclaye, Sylvie
Efremov, Rouslan G.
Hallet, Bernard
Alsteens, David
author_role author
author2 Shkumatov, Alexander V.
Liu, Yun
Stulemeijer, Claire
Derclaye, Sylvie
Efremov, Rouslan G.
Hallet, Bernard
Alsteens, David
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv TRANSPOSITION
PROTEIN-DNA INTERACTION
DYNAMIC FORCE SPECTROSCOPY
AFM
topic TRANSPOSITION
PROTEIN-DNA INTERACTION
DYNAMIC FORCE SPECTROSCOPY
AFM
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Transposon Tn4430 belongs to a widespread family of bacterial transposons, the Tn3 family, whichplays a prevalent role in the dissemination of antibiotic resistance among pathogens. Despite recentdata on the structural architecture of the transposition complex, the molecular mechanisms underlyingthe replicative transposition of these elements arestill poorly understood. Here, we use force-distancecurve-based atomic force microscopy to probe thebinding of the TnpA transposase of Tn4430 to DNAmolecules containing one or two transposon endsand to extract the thermodynamic and kinetic parameters of transposition complex assembly. Comparing wild-type TnpA with previously isolated deregulated TnpA mutants supports a stepwise pathwayfor transposition complex formation and activationduring which TnpA first binds as a dimer to a single transposon end and then undergoes a structuraltransition that enables it to bind the second end cooperatively and to become activated for transpositioncatalysis, the latter step occurring at a much fasterrate for the TnpA mutants. Our study thus providesan unprecedented approach to probe the dynamic ofa complex DNA processing machinery at the singleparticle level.
Fil: Fernandez, Maricruz. Université Catholique de Louvain; Bélgica. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigaciones en Tecnologías Energéticas y Materiales Avanzados. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Tecnologías Energéticas y Materiales Avanzados; Argentina
Fil: Shkumatov, Alexander V.. Faculty Of Sciences And Bioengineering Sciences ; Vrije Universiteit Brussel;
Fil: Liu, Yun. Faculty Of Sciences And Bioengineering Sciences ; Vrije Universiteit Brussel;
Fil: Stulemeijer, Claire. Université Catholique de Louvain; Bélgica
Fil: Derclaye, Sylvie. Université Catholique de Louvain; Bélgica
Fil: Efremov, Rouslan G.. Faculty Of Sciences And Bioengineering Sciences ; Vrije Universiteit Brussel;
Fil: Hallet, Bernard. Université Catholique de Louvain; Bélgica
Fil: Alsteens, David. Université Catholique de Louvain; Bélgica
description Transposon Tn4430 belongs to a widespread family of bacterial transposons, the Tn3 family, whichplays a prevalent role in the dissemination of antibiotic resistance among pathogens. Despite recentdata on the structural architecture of the transposition complex, the molecular mechanisms underlyingthe replicative transposition of these elements arestill poorly understood. Here, we use force-distancecurve-based atomic force microscopy to probe thebinding of the TnpA transposase of Tn4430 to DNAmolecules containing one or two transposon endsand to extract the thermodynamic and kinetic parameters of transposition complex assembly. Comparing wild-type TnpA with previously isolated deregulated TnpA mutants supports a stepwise pathwayfor transposition complex formation and activationduring which TnpA first binds as a dimer to a single transposon end and then undergoes a structuraltransition that enables it to bind the second end cooperatively and to become activated for transpositioncatalysis, the latter step occurring at a much fasterrate for the TnpA mutants. Our study thus providesan unprecedented approach to probe the dynamic ofa complex DNA processing machinery at the singleparticle level.
publishDate 2023
dc.date.none.fl_str_mv 2023-04
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/11336/229930
Fernandez, Maricruz; Shkumatov, Alexander V.; Liu, Yun; Stulemeijer, Claire; Derclaye, Sylvie; et al.; AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements; Oxford University Press; Nucleic Acids Research; 51; 10; 4-2023; 4929-4941
0305-1048
1362-4962
CONICET Digital
CONICET
url http://hdl.handle.net/11336/229930
identifier_str_mv Fernandez, Maricruz; Shkumatov, Alexander V.; Liu, Yun; Stulemeijer, Claire; Derclaye, Sylvie; et al.; AFM-based force spectroscopy unravels stepwise formation of the DNA transposition complex in the widespread Tn3 family mobile genetic elements; Oxford University Press; Nucleic Acids Research; 51; 10; 4-2023; 4929-4941
0305-1048
1362-4962
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://academic.oup.com/nar/advance-article/doi/10.1093/nar/gkad241/7110757
info:eu-repo/semantics/altIdentifier/doi/10.1093/nar/gkad241
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
dc.publisher.none.fl_str_mv Oxford University Press
publisher.none.fl_str_mv Oxford University Press
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
_version_ 1844614239180816384
score 13.070432

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