Identification of a miniature Sae2/Ctp1/CtIP ortholog from Paramecium tetraurelia required for sexual reproduction and DNA double-strand break repair
| Autor: | Antoine Marmignon, Julia Godau, Anika Trenner, Christine von Aesch, Emeline Dubois, Aurélie Kapusta, Mireille Bétermier, Lauriane Simon, Lorenza P. Ferretti, Alessandro A. Sartori, Raphael Guerois |
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| Přispěvatelé: | Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Génétique, Reproduction et Développement (GReD ), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre National de la Recherche Scientifique (CNRS), Department of Human Genetics [Salt Lake City], University of Utah, Assemblage moléculaire et intégrité du génome (AMIG), Département Biochimie, Biophysique et Biologie Structurale (B3S), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Réarrangements programmés du génome (MICMAC), Département Biologie des Génomes (DBG), University of Zurich, Bétermier, Mireille, Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]) |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
1303 Biochemistry
DNA Repair sae2 [SDV]Life Sciences [q-bio] Mutant Amino Acid Motifs Protozoan Proteins Biochemistry endonuclease 1307 Cell Biology Endonuclease chemistry.chemical_compound 0302 clinical medicine DNA Breaks Double-Stranded Conserved Sequence 0303 health sciences damage response biology Reproduction 10061 Institute of Molecular Cancer Research rad32(mre11) nuclease Cell biology Meiosis mre11 complex 030220 oncology & carcinogenesis DNA end resection 610 Medicine & health 03 medical and health sciences Mre11 complex 1312 Molecular Biology DNA double-strand breaks Amino Acid Sequence Homologous recombination gene Molecular Biology Gene 030304 developmental biology Nuclease Sequence Homology Amino Acid Cell Biology DNA Protozoan human ctip chemistry CtIP end-resection biology.protein 570 Life sciences ctp1 Paramecium tetraurelia protein DNA |
| Zdroj: | DNA Repair DNA Repair, 2019, 77, pp.96--108. ⟨10.1016/j.dnarep.2019.03.011⟩ |
| Popis: | International audience; DNA double-strand breaks (DSBs) induced by genotoxic agents can cause cell death or contribute to chromosomal instability, a major driving force of cancer. By contrast, Spo11-dependent DSBs formed during meiosis are aimed at generating genetic diversity. In eukaryotes, CtIP and the Mre11 nuclease complex are essential for accurate processing and repair of both unscheduled and programmed DSBs by homologous recombination (HR). Here, we applied bioinformatics and genetic analysis to identify Paramecium tetraurelia CtIP (PtCtIP), the smallest known Sae2/Ctp1/CtIP ortholog, as a key factor for the completion of meiosis and the recovery of viable sexual progeny. Using in vitro assays, we find that purified recombinant PtCtIP preferentially binds to double-stranded DNA substrates but does not contain intrinsic nuclease activity. Moreover, mutation of the evolutionarily conserved C-terminal 'RHR' motif abrogates DNA binding of PtCtIP but not its ability to functionally interact with Mre11. Translating our findings into mammalian cells, we provide evidence that disruption of the 'RHR' motif abrogates accumulation of human CtIP at sites of DSBs. Consequently, cells expressing the DNA binding mutant CtIPR837A/R839A are defective in DSB resection and HR. Collectively, our work highlights minimal structural requirements for CtIP protein family members to facilitate the processing of DSBs, thereby maintaining genome stability as well as enabling sexual reproduction. |
| Databáze: | OpenAIRE |
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