Régulation de la réponse à divers stress et réparation des cassures double brin de l’ADN chez la bactérie Deinococcus radiodurans

Abstract : The Deinococcus radiodurans bacterium exhibits resistance to γ and UV radiation, desiccation and oxidative stress. The molecular mechanisms contributing to the radioresistance of D. radiodurans include very efficient DNA repair mechanisms and ROS detoxification systems, protein protection against oxidation, a compact nucleoid structure and a subset of Deinococcus specific genes which are strongly induced after γ radiation. The ddrI (DNA damage response) gene is highly up-regulated after exposure to γ radiation and encodes a transcription factor belonging to the CRP (cAMP receptor protein) family. Compared to wild type cells, cells devoid of DdrI display defects in cell division and/or DNA segregation and is sensitive to DNA damaging agents, oxidative stress and heat shock treatment. In silico predictions of putative DdrI targets suggest that hundreds of genes,belonging to various cellular processes (DNA replication and repair, oxidative stress and heat shock responses, regulation of transcription and signal transduction) may be regulated by DdrI. The pseudopalindromic 5’TGTGA(N6)TCACA3’ consensus sequence, extrapolated from 115 potential DdrI binding sites, is specifically bound by DdrI only in presence of cAMP. After heat shock treatment, DdrI is involved directly or indirectly, in the induction of heat shock response genes coding proteases, proteins involved in DNA, lipid, carbohydrate metabolism and a translation inhibitor. Among the Deinococcus specific proteins required for radioresistance, the PprA protein was shown to play a major role for accurate chromosome segregation and cell division after completion of DNA repair. Here, we analyzed the cellular role of the RecN protein belonging to the SMC family and, surprisingly, observed that the absence of the RecN protein suppressed the sensitivity of cells devoid of the PprA protein to γ- and UV-irradiation and to treatment with mitomycin C or DNA gyrase inhibitors. The absence of RecN also alleviated the DNA segregation defects displayed by the ΔpprA cells recovering from irradiation. After irradiation, the absence of RecN reduced recombination between chromosomal and plasmid DNA, indicating that the RecN protein is important for recombinational repair of DNA lesions. Here, we propose a model in which RecN, by favoring recombinational repair of DNA double strand breaks, requires the PprA protein to facilitate the recruitment of the DNA topoisomerases to resolve the topological constraints generated by DNA double strand break repair through homologous recombination.
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Laura Meyer. Régulation de la réponse à divers stress et réparation des cassures double brin de l’ADN chez la bactérie Deinococcus radiodurans. Biochimie, Biologie Moléculaire. Université Paris-Saclay, 2018. Français. ⟨NNT : 2018SACLS422⟩. ⟨tel-02145221v2⟩



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