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Role of Homologous Recombination Genes in Repair of Alkylation Base Damage by Candida albicans

Candida albicans mutants deficient in homologous recombination (HR) are extremely sensitive to the alkylating agent methyl-methane-sulfonate (MMS). Here, we have investigated the role of HR genes in the protection and repair of C. albicans chromosomes by taking advantage of the heat-labile property...

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Autores principales: Ciudad, Toni, Bellido, Alberto, Andaluz, Encarnación, Hermosa, Belén, Larriba, Germán
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6162806/
https://www.ncbi.nlm.nih.gov/pubmed/30205450
http://dx.doi.org/10.3390/genes9090447
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author Ciudad, Toni
Bellido, Alberto
Andaluz, Encarnación
Hermosa, Belén
Larriba, Germán
author_facet Ciudad, Toni
Bellido, Alberto
Andaluz, Encarnación
Hermosa, Belén
Larriba, Germán
author_sort Ciudad, Toni
collection PubMed
description Candida albicans mutants deficient in homologous recombination (HR) are extremely sensitive to the alkylating agent methyl-methane-sulfonate (MMS). Here, we have investigated the role of HR genes in the protection and repair of C. albicans chromosomes by taking advantage of the heat-labile property (55 °C) of MMS-induced base damage. Acute MMS treatments of cycling cells caused chromosome fragmentation in vitro (55 °C) due to the generation of heat-dependent breaks (HDBs), but not in vivo (30 °C). Following removal of MMS wild type, cells regained the chromosome ladder regardless of whether they were transferred to yeast extract/peptone/dextrose (YPD) or to phosphate buffer saline (PBS); however, repair of HDB/chromosome restitution was faster in YPD, suggesting that it was accelerated by metabolic energy and further fueled by the subsequent overgrowth of survivors. Compared to wild type CAI4, chromosome restitution in YPD was not altered in a Carad59 isogenic derivative, whereas it was significantly delayed in Carad51 and Carad52 counterparts. However, when post-MMS incubation took place in PBS, chromosome restitution in wild type and HR mutants occurred with similar kinetics, suggesting that the exquisite sensitivity of Carad51 and Carad52 mutants to MMS is due to defective fork restart. Overall, our results demonstrate that repair of HDBs by resting cells of C. albicans is rather independent of CaRad51, CaRad52, and CaRad59, suggesting that it occurs mainly by base excision repair (BER).
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spelling pubmed-61628062018-10-10 Role of Homologous Recombination Genes in Repair of Alkylation Base Damage by Candida albicans Ciudad, Toni Bellido, Alberto Andaluz, Encarnación Hermosa, Belén Larriba, Germán Genes (Basel) Article Candida albicans mutants deficient in homologous recombination (HR) are extremely sensitive to the alkylating agent methyl-methane-sulfonate (MMS). Here, we have investigated the role of HR genes in the protection and repair of C. albicans chromosomes by taking advantage of the heat-labile property (55 °C) of MMS-induced base damage. Acute MMS treatments of cycling cells caused chromosome fragmentation in vitro (55 °C) due to the generation of heat-dependent breaks (HDBs), but not in vivo (30 °C). Following removal of MMS wild type, cells regained the chromosome ladder regardless of whether they were transferred to yeast extract/peptone/dextrose (YPD) or to phosphate buffer saline (PBS); however, repair of HDB/chromosome restitution was faster in YPD, suggesting that it was accelerated by metabolic energy and further fueled by the subsequent overgrowth of survivors. Compared to wild type CAI4, chromosome restitution in YPD was not altered in a Carad59 isogenic derivative, whereas it was significantly delayed in Carad51 and Carad52 counterparts. However, when post-MMS incubation took place in PBS, chromosome restitution in wild type and HR mutants occurred with similar kinetics, suggesting that the exquisite sensitivity of Carad51 and Carad52 mutants to MMS is due to defective fork restart. Overall, our results demonstrate that repair of HDBs by resting cells of C. albicans is rather independent of CaRad51, CaRad52, and CaRad59, suggesting that it occurs mainly by base excision repair (BER). MDPI 2018-09-07 /pmc/articles/PMC6162806/ /pubmed/30205450 http://dx.doi.org/10.3390/genes9090447 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ciudad, Toni
Bellido, Alberto
Andaluz, Encarnación
Hermosa, Belén
Larriba, Germán
Role of Homologous Recombination Genes in Repair of Alkylation Base Damage by Candida albicans
title Role of Homologous Recombination Genes in Repair of Alkylation Base Damage by Candida albicans
title_full Role of Homologous Recombination Genes in Repair of Alkylation Base Damage by Candida albicans
title_fullStr Role of Homologous Recombination Genes in Repair of Alkylation Base Damage by Candida albicans
title_full_unstemmed Role of Homologous Recombination Genes in Repair of Alkylation Base Damage by Candida albicans
title_short Role of Homologous Recombination Genes in Repair of Alkylation Base Damage by Candida albicans
title_sort role of homologous recombination genes in repair of alkylation base damage by candida albicans
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6162806/
https://www.ncbi.nlm.nih.gov/pubmed/30205450
http://dx.doi.org/10.3390/genes9090447
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