Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation

Autores
de Tullio, Luisina; Kaniecki, Kyle; Kwon, Youngho; Crickard, J. Brooks; Sung, Patrick; Greene, Eric C.
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Srs2 is a super-family 1 helicase that promotes genome stability by dismantling toxic DNA recombination intermediates. However, the mechanisms by which Srs2 remodels or resolves recombination intermediates remain poorly understood. Here, single-molecule imaging is used to visualize Srs2 in real time as it acts on single-stranded DNA (ssDNA) bound by protein factors that function in recombination. We demonstrate that Srs2 is highly processive and translocates rapidly (∼170 nt per second) in the 3′→5′ direction along ssDNA saturated with replication protein A (RPA). We show that RPA is evicted from DNA during the passage of Srs2. Remarkably, Srs2 also readily removes the recombination mediator Rad52 from RPA-ssDNA and, in doing so, promotes rapid redistribution of both Rad52 and RPA. These findings have important mechanistic implications for understanding how Srs2 and related nucleic acid motor proteins resolve potentially pathogenic nucleoprotein intermediates. De Tullio et al. develop a single-molecule assay for directly visualizing the behavior of the yeast helicase Srs2 as it acts upon single-stranded DNA (ssDNA). These experiments demonstrate that Srs2 is capable of rapid and processive translocation on ssDNA coated with the homologous recombination accessory proteins RPA and Rad52.
Fil: de Tullio, Luisina. Columbia University In The City Of New York; . Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Kaniecki, Kyle. Columbia University; Estados Unidos
Fil: Kwon, Youngho. University of Yale. School of Medicine; Estados Unidos
Fil: Crickard, J. Brooks. Columbia University; Estados Unidos
Fil: Sung, Patrick. University of Yale. School of Medicine; Estados Unidos
Fil: Greene, Eric C.. Columbia University; Estados Unidos
Materia
Dna Curtain
Homologous Recombination
Rad52
Replication Protein A
Single Molecule
Srs2
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:11336/57542
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/57542
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulationde Tullio, LuisinaKaniecki, KyleKwon, YounghoCrickard, J. BrooksSung, PatrickGreene, Eric C.Dna CurtainHomologous RecombinationRad52Replication Protein ASingle MoleculeSrs2https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Srs2 is a super-family 1 helicase that promotes genome stability by dismantling toxic DNA recombination intermediates. However, the mechanisms by which Srs2 remodels or resolves recombination intermediates remain poorly understood. Here, single-molecule imaging is used to visualize Srs2 in real time as it acts on single-stranded DNA (ssDNA) bound by protein factors that function in recombination. We demonstrate that Srs2 is highly processive and translocates rapidly (∼170 nt per second) in the 3′→5′ direction along ssDNA saturated with replication protein A (RPA). We show that RPA is evicted from DNA during the passage of Srs2. Remarkably, Srs2 also readily removes the recombination mediator Rad52 from RPA-ssDNA and, in doing so, promotes rapid redistribution of both Rad52 and RPA. These findings have important mechanistic implications for understanding how Srs2 and related nucleic acid motor proteins resolve potentially pathogenic nucleoprotein intermediates. De Tullio et al. develop a single-molecule assay for directly visualizing the behavior of the yeast helicase Srs2 as it acts upon single-stranded DNA (ssDNA). These experiments demonstrate that Srs2 is capable of rapid and processive translocation on ssDNA coated with the homologous recombination accessory proteins RPA and Rad52.Fil: de Tullio, Luisina. Columbia University In The City Of New York; . Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaFil: Kaniecki, Kyle. Columbia University; Estados UnidosFil: Kwon, Youngho. University of Yale. School of Medicine; Estados UnidosFil: Crickard, J. Brooks. Columbia University; Estados UnidosFil: Sung, Patrick. University of Yale. School of Medicine; Estados UnidosFil: Greene, Eric C.. Columbia University; Estados UnidosElsevier B.V.2017-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/57542de Tullio, Luisina; Kaniecki, Kyle; Kwon, Youngho; Crickard, J. Brooks; Sung, Patrick; et al.; Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation; Elsevier B.V.; Cell Reports; 21; 3; 10-2017; 570-5772211-12472211-1247CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pubmed/29045827info:eu-repo/semantics/altIdentifier/doi/10.1016/j.celrep.2017.09.073info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:28:35Zoai:ri.conicet.gov.ar:11336/57542instacron: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 10:28:35.557CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation
title Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation
spellingShingle Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation
de Tullio, Luisina
Dna Curtain
Homologous Recombination
Rad52
Replication Protein A
Single Molecule
Srs2
title_short Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation
title_full Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation
title_fullStr Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation
title_full_unstemmed Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation
title_sort Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation
dc.creator.none.fl_str_mv de Tullio, Luisina
Kaniecki, Kyle
Kwon, Youngho
Crickard, J. Brooks
Sung, Patrick
Greene, Eric C.
author de Tullio, Luisina
author_facet de Tullio, Luisina
Kaniecki, Kyle
Kwon, Youngho
Crickard, J. Brooks
Sung, Patrick
Greene, Eric C.
author_role author
author2 Kaniecki, Kyle
Kwon, Youngho
Crickard, J. Brooks
Sung, Patrick
Greene, Eric C.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Dna Curtain
Homologous Recombination
Rad52
Replication Protein A
Single Molecule
Srs2
topic Dna Curtain
Homologous Recombination
Rad52
Replication Protein A
Single Molecule
Srs2
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Srs2 is a super-family 1 helicase that promotes genome stability by dismantling toxic DNA recombination intermediates. However, the mechanisms by which Srs2 remodels or resolves recombination intermediates remain poorly understood. Here, single-molecule imaging is used to visualize Srs2 in real time as it acts on single-stranded DNA (ssDNA) bound by protein factors that function in recombination. We demonstrate that Srs2 is highly processive and translocates rapidly (∼170 nt per second) in the 3′→5′ direction along ssDNA saturated with replication protein A (RPA). We show that RPA is evicted from DNA during the passage of Srs2. Remarkably, Srs2 also readily removes the recombination mediator Rad52 from RPA-ssDNA and, in doing so, promotes rapid redistribution of both Rad52 and RPA. These findings have important mechanistic implications for understanding how Srs2 and related nucleic acid motor proteins resolve potentially pathogenic nucleoprotein intermediates. De Tullio et al. develop a single-molecule assay for directly visualizing the behavior of the yeast helicase Srs2 as it acts upon single-stranded DNA (ssDNA). These experiments demonstrate that Srs2 is capable of rapid and processive translocation on ssDNA coated with the homologous recombination accessory proteins RPA and Rad52.
Fil: de Tullio, Luisina. Columbia University In The City Of New York; . Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Kaniecki, Kyle. Columbia University; Estados Unidos
Fil: Kwon, Youngho. University of Yale. School of Medicine; Estados Unidos
Fil: Crickard, J. Brooks. Columbia University; Estados Unidos
Fil: Sung, Patrick. University of Yale. School of Medicine; Estados Unidos
Fil: Greene, Eric C.. Columbia University; Estados Unidos
description Srs2 is a super-family 1 helicase that promotes genome stability by dismantling toxic DNA recombination intermediates. However, the mechanisms by which Srs2 remodels or resolves recombination intermediates remain poorly understood. Here, single-molecule imaging is used to visualize Srs2 in real time as it acts on single-stranded DNA (ssDNA) bound by protein factors that function in recombination. We demonstrate that Srs2 is highly processive and translocates rapidly (∼170 nt per second) in the 3′→5′ direction along ssDNA saturated with replication protein A (RPA). We show that RPA is evicted from DNA during the passage of Srs2. Remarkably, Srs2 also readily removes the recombination mediator Rad52 from RPA-ssDNA and, in doing so, promotes rapid redistribution of both Rad52 and RPA. These findings have important mechanistic implications for understanding how Srs2 and related nucleic acid motor proteins resolve potentially pathogenic nucleoprotein intermediates. De Tullio et al. develop a single-molecule assay for directly visualizing the behavior of the yeast helicase Srs2 as it acts upon single-stranded DNA (ssDNA). These experiments demonstrate that Srs2 is capable of rapid and processive translocation on ssDNA coated with the homologous recombination accessory proteins RPA and Rad52.
publishDate 2017
dc.date.none.fl_str_mv 2017-10
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/11336/57542
de Tullio, Luisina; Kaniecki, Kyle; Kwon, Youngho; Crickard, J. Brooks; Sung, Patrick; et al.; Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation; Elsevier B.V.; Cell Reports; 21; 3; 10-2017; 570-577
2211-1247
2211-1247
CONICET Digital
CONICET
url http://hdl.handle.net/11336/57542
identifier_str_mv de Tullio, Luisina; Kaniecki, Kyle; Kwon, Youngho; Crickard, J. Brooks; Sung, Patrick; et al.; Yeast Srs2 helicase promotes redistribution of single-stranded DNA-Bound RPA and Rad52 in homologous recombination regulation; Elsevier B.V.; Cell Reports; 21; 3; 10-2017; 570-577
2211-1247
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://www.ncbi.nlm.nih.gov/pubmed/29045827
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.celrep.2017.09.073
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 Elsevier B.V.
publisher.none.fl_str_mv Elsevier B.V.
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 13.070432

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