Impact of Pif1 translational mechanism on genetic interactions with YEN1ON and other DNA repair enzymes

Homologous recombination (HR) entails the formation of several branched recombination intermediates that must be timely disengaged to safeguard chromosome segregation and cell viability. Hence, cells are endowed with DNA helicases and structure-selective endonucleases (SSEs) that sever these physica...

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Detalles Bibliográficos
Autor: Lama Díaz, Tomás
Tipo de recurso: tesis doctoral
Fecha de publicación:2023
País:España
Institución:Universidad de Santiago de Compostela (USC)
Repositorio:Minerva. Repositorio Institucional de la Universidad de Santiago de Compostela
Idioma:inglés
OAI Identifier:oai:minerva.usc.gal:10347/30882
Acceso en línea:http://hdl.handle.net/10347/30882
Access Level:acceso abierto
Palabra clave:230221 Biología molecular
240902 Ingeniería genética
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spelling Impact of Pif1 translational mechanism on genetic interactions with YEN1ON and other DNA repair enzymesLama Díaz, Tomás230221 Biología molecular240902 Ingeniería genéticaHomologous recombination (HR) entails the formation of several branched recombination intermediates that must be timely disengaged to safeguard chromosome segregation and cell viability. Hence, cells are endowed with DNA helicases and structure-selective endonucleases (SSEs) that sever these physical connections between DNA joint molecules prior to cell division. Surprinsingly, the activation of SSEs is delayed until the final stages of cell cycle. This strategy, conserved from yeast to humans, suggests that this tight control could be crucial to prevent the unscheduled processing of DNA replication and repair intermediates. To understand the biological relevance of this strict regulatory system, we searched for genetic interactions between a constitutively active version of the YEN1 nuclease (YEN1-ON) and key helicases involved in DNA replication and repair. Here, we show that deletion of the conserved PIF1 helicase in a YEN1-ON strain results in a dramatic reduction of its viability under genotoxic stress. This suggests that the unscheduled nucleolytic processing of secondary DNA structures accumulated in the absence of PIF1 is detrimental for cells. PIF1 encodes both mitochondrial and nuclear isoforms of the enzyme, but widely employed separation-of-function alleles (nuclear or mitochondrial-specific) failed to recapitulate such genetic interaction. This prompted us to delve into the translational mechanism of Pif1, leading us to the refinement of the molecular mechanism of alternative translation initiation for PIF1 mRNA, the discovery of a new nuclear Pif1 isoform and the development of the first bona-fide nuclear-null Pif1 isoform.González Blanco, MiguelUniversidade de Santiago de Compostela. Escola de Doutoramento Internacional (EDIUS)20232023-01-0120232023-01-01doctoral thesishttp://purl.org/coar/resource_type/c_db06info:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/10347/30882reponame:Minerva. Repositorio Institucional de la Universidad de Santiago de Compostelainstname:Universidad de Santiago de Compostela (USC)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution-NonCommercial-NoDerivatives 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:minerva.usc.gal:10347/308822026-06-15T12:47:27Z
dc.title.none.fl_str_mv Impact of Pif1 translational mechanism on genetic interactions with YEN1ON and other DNA repair enzymes
title Impact of Pif1 translational mechanism on genetic interactions with YEN1ON and other DNA repair enzymes
spellingShingle Impact of Pif1 translational mechanism on genetic interactions with YEN1ON and other DNA repair enzymes
Lama Díaz, Tomás
230221 Biología molecular
240902 Ingeniería genética
title_short Impact of Pif1 translational mechanism on genetic interactions with YEN1ON and other DNA repair enzymes
title_full Impact of Pif1 translational mechanism on genetic interactions with YEN1ON and other DNA repair enzymes
title_fullStr Impact of Pif1 translational mechanism on genetic interactions with YEN1ON and other DNA repair enzymes
title_full_unstemmed Impact of Pif1 translational mechanism on genetic interactions with YEN1ON and other DNA repair enzymes
title_sort Impact of Pif1 translational mechanism on genetic interactions with YEN1ON and other DNA repair enzymes
dc.creator.none.fl_str_mv Lama Díaz, Tomás
author Lama Díaz, Tomás
author_facet Lama Díaz, Tomás
author_role author
dc.contributor.none.fl_str_mv González Blanco, Miguel
Universidade de Santiago de Compostela. Escola de Doutoramento Internacional (EDIUS)
dc.subject.none.fl_str_mv 230221 Biología molecular
240902 Ingeniería genética
topic 230221 Biología molecular
240902 Ingeniería genética
description Homologous recombination (HR) entails the formation of several branched recombination intermediates that must be timely disengaged to safeguard chromosome segregation and cell viability. Hence, cells are endowed with DNA helicases and structure-selective endonucleases (SSEs) that sever these physical connections between DNA joint molecules prior to cell division. Surprinsingly, the activation of SSEs is delayed until the final stages of cell cycle. This strategy, conserved from yeast to humans, suggests that this tight control could be crucial to prevent the unscheduled processing of DNA replication and repair intermediates. To understand the biological relevance of this strict regulatory system, we searched for genetic interactions between a constitutively active version of the YEN1 nuclease (YEN1-ON) and key helicases involved in DNA replication and repair. Here, we show that deletion of the conserved PIF1 helicase in a YEN1-ON strain results in a dramatic reduction of its viability under genotoxic stress. This suggests that the unscheduled nucleolytic processing of secondary DNA structures accumulated in the absence of PIF1 is detrimental for cells. PIF1 encodes both mitochondrial and nuclear isoforms of the enzyme, but widely employed separation-of-function alleles (nuclear or mitochondrial-specific) failed to recapitulate such genetic interaction. This prompted us to delve into the translational mechanism of Pif1, leading us to the refinement of the molecular mechanism of alternative translation initiation for PIF1 mRNA, the discovery of a new nuclear Pif1 isoform and the development of the first bona-fide nuclear-null Pif1 isoform.
publishDate 2023
dc.date.none.fl_str_mv 2023
2023-01-01
2023
2023-01-01
dc.type.none.fl_str_mv doctoral thesis
http://purl.org/coar/resource_type/c_db06
dc.type.openaire.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
dc.identifier.none.fl_str_mv http://hdl.handle.net/10347/30882
url http://hdl.handle.net/10347/30882
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Minerva. Repositorio Institucional de la Universidad de Santiago de Compostela
instname:Universidad de Santiago de Compostela (USC)
instname_str Universidad de Santiago de Compostela (USC)
reponame_str Minerva. Repositorio Institucional de la Universidad de Santiago de Compostela
collection Minerva. Repositorio Institucional de la Universidad de Santiago de Compostela
repository.name.fl_str_mv
repository.mail.fl_str_mv
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