Role of DisA and c-di-AMP in the DNA damage response of exponentially growing "Bacillus subtilis" cells

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid. Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 10-06-2015

Detalles Bibliográficos
Autor: Gándara, Carolina
Tipo de recurso: tesis doctoral
Fecha de publicación:2015
País:España
Institución:Universidad Autónoma de Madrid
Repositorio:Biblos-e Archivo. Repositorio Institucional de la UAM
Idioma:español
inglés
OAI Identifier:oai:repositorio.uam.es:10486/667520
Acceso en línea:http://hdl.handle.net/10486/667520
Access Level:acceso abierto
Palabra clave:Bacilus subtilis - Tesis doctorales
Biología y Biomedicina / Biología
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spelling Role of DisA and c-di-AMP in the DNA damage response of exponentially growing "Bacillus subtilis" cellsGándara, CarolinaBacilus subtilis - Tesis doctoralesBiología y Biomedicina / BiologíaTesis doctoral inédita leída en la Universidad Autónoma de Madrid. Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 10-06-2015Cells are constantly subjected to DNA lesions, which can result in replication fork stalling or collapse. A variety of DNA repair pathways have evolved to respond to the different types of exogenous and endogenous DNA damaging agents. A complex regulation exists to coordinate these processes, which are called DNA damage response (DDRs). DisA was initially described as a DNA damage checkpoint protein that ensures the genome integrity of Bacillus subtilis spores. However, basal levels of DisA were detected in vegetative cell extracts and highly dynamic foci were observed in vegetative cells expressing DisA-YFP under its native promoter. Since DisA is also present in non-spore forming bacteria, a more general role for DisA, that is not restricted to sporulation, was proposed. In this work, it was demonstrated that c-di-AMP and the diadenilate cyclases (DAC), DisA and CdaA, contribute to elicit repair of stalled and collapsed forks, in exponentially growing B. subtilis cells. Absence of DisA or decreased intracellular levels of c-di-AMP impaired survival to alkylation DNA damage induced by methyl methanesulfonate (MMS). On the other hand, CdaA is involved in oxidative damage repair. The radA and disA functions were epistatic for MMS and showed also epistasis with functions involved in Holliday junction (HJ) processing. Both radA and disA were epistatic with recG and recU, while non-epistatic with ruvAB. In vivo, DisA-YFP foci lost its dynamics in the presence of unresolved HJs (ΔrecU context). Purified DisA showed DAC activity with preference for ATP over dATP to produce c-di- AMP. DisA could bind different DNA structures without specificity, although only highly stacked HJs could modulate DisA c-di-AMP synthesis. For the first time it was demonstrated that RadA/Sms could bind DNA, in the absence or presence of ATP, and with more affinity for single-stranded (ss) DNA and HJs. In vitro assays showed that DisA strongly inhibits RecG and RuvAB helicase activities. DisA also negatively affects RecU resolvase activity and RecAmediated strand exchange reactions. Altogether, these results indicate that, in stalled forks induced by the presence of alkyl adducts, DisA modulates the DDR by binding branched DNA and reducing the synthesis of c-di-AMP. More precisely, DisA modulates recombinational DNA repair by protecting branched recombination intermediates, and therefore preventing fork reversal, HJ resolution and strand exchange. It is likely that DisA stimulate other specific DNA repair pathways, such as BER, and favours TLS since slightly lower mutation frequencies were detected in the absence of DisA.Alonso, Juan CarlosDepartamento de Biología MolecularFacultad de CienciasCSIC. Centro Nacional de Biotecnología (CNB)20152015-06-10doctoral thesishttp://purl.org/coar/resource_type/c_db06NAhttp://purl.org/coar/version/c_be7fb7dd8ff6fe43info:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/10486/667520reponame:Biblos-e Archivo. Repositorio Institucional de la UAMinstname:Universidad Autónoma de MadridEspañolspaInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:repositorio.uam.es:10486/6675202026-06-23T12:46:27Z
dc.title.none.fl_str_mv Role of DisA and c-di-AMP in the DNA damage response of exponentially growing "Bacillus subtilis" cells
title Role of DisA and c-di-AMP in the DNA damage response of exponentially growing "Bacillus subtilis" cells
spellingShingle Role of DisA and c-di-AMP in the DNA damage response of exponentially growing "Bacillus subtilis" cells
Gándara, Carolina
Bacilus subtilis - Tesis doctorales
Biología y Biomedicina / Biología
title_short Role of DisA and c-di-AMP in the DNA damage response of exponentially growing "Bacillus subtilis" cells
title_full Role of DisA and c-di-AMP in the DNA damage response of exponentially growing "Bacillus subtilis" cells
title_fullStr Role of DisA and c-di-AMP in the DNA damage response of exponentially growing "Bacillus subtilis" cells
title_full_unstemmed Role of DisA and c-di-AMP in the DNA damage response of exponentially growing "Bacillus subtilis" cells
title_sort Role of DisA and c-di-AMP in the DNA damage response of exponentially growing "Bacillus subtilis" cells
dc.creator.none.fl_str_mv Gándara, Carolina
author Gándara, Carolina
author_facet Gándara, Carolina
author_role author
dc.contributor.none.fl_str_mv Alonso, Juan Carlos
Departamento de Biología Molecular
Facultad de Ciencias
CSIC. Centro Nacional de Biotecnología (CNB)
dc.subject.none.fl_str_mv Bacilus subtilis - Tesis doctorales
Biología y Biomedicina / Biología
topic Bacilus subtilis - Tesis doctorales
Biología y Biomedicina / Biología
description Tesis doctoral inédita leída en la Universidad Autónoma de Madrid. Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 10-06-2015
publishDate 2015
dc.date.none.fl_str_mv 2015
2015-06-10
dc.type.none.fl_str_mv doctoral thesis
http://purl.org/coar/resource_type/c_db06
NA
http://purl.org/coar/version/c_be7fb7dd8ff6fe43
dc.type.openaire.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
dc.identifier.none.fl_str_mv http://hdl.handle.net/10486/667520
url http://hdl.handle.net/10486/667520
dc.language.none.fl_str_mv Español
spa
Inglés
eng
language_invalid_str_mv Español
Inglés
language spa
eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
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
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Biblos-e Archivo. Repositorio Institucional de la UAM
instname:Universidad Autónoma de Madrid
instname_str Universidad Autónoma de Madrid
reponame_str Biblos-e Archivo. Repositorio Institucional de la UAM
collection Biblos-e Archivo. Repositorio Institucional de la UAM
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,300719

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