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Genes 2018, 9(9), 447; https://doi.org/10.3390/genes9090447

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

Departamento de Microbiología, Facultad de Ciencias, Universidad de Extremadura, 06071 Badajoz, Spain
These authors contributed equally to this work.
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Received: 14 August 2018 / Accepted: 27 August 2018 / Published: 7 September 2018
(This article belongs to the Special Issue Fungal Pathogenesis in Humans: The Growing Threat)
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Abstract

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). View Full-Text
Keywords: homologous recombination; Candida albicans; alkylation damage; repair homologous recombination; Candida albicans; alkylation damage; repair
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Ciudad, T.; Bellido, A.; Andaluz, E.; Hermosa, B.; Larriba, G. Role of Homologous Recombination Genes in Repair of Alkylation Base Damage by Candida albicans. Genes 2018, 9, 447.

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