PAXX Is an Accessory c-NHEJ Factor that Associates with Ku70 and Has Overlapping Functions with XLF

In mammalian cells, classical non-homologous end joining (c-NHEJ) is critical for DNA double-strand break repair induced by ionizing radiation and during V(D)J recombination in developing B and T lymphocytes. Recently, PAXX was identified as a c-NHEJ core component. We report here that PAXX-deficien...

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Veröffentlicht in:	Cell reports (Cambridge) Jg. 17; H. 2; S. 541 - 555
Hauptverfasser:	Tadi, Satish K., Tellier-Lebègue, Carine, Nemoz, Clément, Drevet, Pascal, Audebert, Stéphane, Roy, Sunetra, Meek, Katheryn, Charbonnier, Jean-Baptiste, Modesti, Mauro
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	United States Elsevier Inc 04.10.2016 Elsevier
Schlagworte:	B-Lymphocytes - metabolism DNA Breaks, Double-Stranded DNA double-strand break repair DNA End-Joining Repair - genetics DNA ligase 4 DNA Repair - genetics DNA Repair Enzymes - genetics DNA Repair Enzymes - metabolism DNA-Binding Proteins - chemistry DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism HCT116 Cells Humans Ku Autoantigen - chemistry Ku Autoantigen - genetics Ku Autoantigen - metabolism Ku70 Life Sciences MMEJ NHEJ PAXX T-Lymphocytes - metabolism V(D)J recombination V(D)J Recombination - genetics XLF XRCC4 Ku70 DNA double-strand break repair MMEJ XLF PAXX XRCC4 V(D)J recombination DNA ligase 4 Ku NHEJ
ISSN:	2211-1247, 2211-1247
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Abstract	In mammalian cells, classical non-homologous end joining (c-NHEJ) is critical for DNA double-strand break repair induced by ionizing radiation and during V(D)J recombination in developing B and T lymphocytes. Recently, PAXX was identified as a c-NHEJ core component. We report here that PAXX-deficient cells exhibit a cellular phenotype uncharacteristic of a deficiency in c-NHEJ core components. PAXX-deficient cells display normal sensitivity to radiomimetic drugs, are proficient in transient V(D)J recombination assays, and do not shift toward higher micro-homology usage in plasmid repair assays. Although PAXX-deficient cells lack c-NHEJ phenotypes, PAXX forms a stable ternary complex with Ku bound to DNA. Formation of this complex involves an interaction with Ku70 and requires a bare DNA extension for stability. Moreover, the relatively weak Ku-dependent stimulation of LIG4/XRCC4 activity by PAXX is unmasked by XLF ablation. Thus, PAXX plays an accessory role during c-NHEJ that is largely overlapped by XLF’s function. [Display omitted] •PAXX interaction with Ku threaded onto DNA requires a bare DNA extension•PAXX interacts with the Ku70 subunit•PAXX is dispensable for V(D)J recombination and has no core c-NHEJ phenotype•PAXX’s function in c-NHEJ is masked by XLF Tadi et al. find that PAXX interacts with Ku threaded on DNA via the Ku70 subunit and requires a bare DNA extension for stability. They show that PAXX’s function overlaps with c-NHEJ factor XLF, explaining why its ablation has only a mild effect on NHEJ.
AbstractList	In mammalian cells, classical non-homologous end joining (c-NHEJ) is critical for DNA double-strand break repair induced by ionizing radiation and during V(D)J recombination in developing B and T lymphocytes. Recently, PAXX was identified as a c-NHEJ core component. We report here that PAXX-deficient cells exhibit a cellular phenotype uncharacteristic of a deficiency in c-NHEJ core components. PAXX-deficient cells display normal sensitivity to radiomimetic drugs, are proficient in transient V(D)J recombination assays, and do not shift toward higher micro-homology usage in plasmid repair assays. Although PAXX-deficient cells lack c-NHEJ phenotypes, PAXX forms a stable ternary complex with Ku bound to DNA. Formation of this complex involves an interaction with Ku70 and requires a bare DNA extension for stability. Moreover, the relatively weak Ku-dependent stimulation of LIG4/XRCC4 activity by PAXX is unmasked by XLF ablation. Thus, PAXX plays an accessory role during c-NHEJ that is largely overlapped by XLF's function. In mammalian cells, classical non-homologous end joining (c-NHEJ) is critical for DNA double-strand break repair induced by ionizing radiation and during V(D)J recombination in developing B and T lymphocytes. Recently, PAXX was identified as a c-NHEJ core component. We report here that PAXX-deficient cells exhibit a cellular phenotype uncharacteristic of a deficiency in c-NHEJ core components. PAXX-deficient cells display normal sensitivity to radiomimetic drugs, are proficient in transient V(D)J recombination assays, and do not shift toward higher micro-homology usage in plasmid repair assays. Although PAXX-deficient cells lack c-NHEJ phenotypes, PAXX forms a stable ternary complex with Ku bound to DNA. Formation of this complex involves an interaction with Ku70 and requires a bare DNA extension for stability. Moreover, the relatively weak Ku-dependent stimulation of LIG4/XRCC4 activity by PAXX is unmasked by XLF ablation. Thus, PAXX plays an accessory role during c-NHEJ that is largely overlapped by XLF’s function. [Display omitted] •PAXX interaction with Ku threaded onto DNA requires a bare DNA extension•PAXX interacts with the Ku70 subunit•PAXX is dispensable for V(D)J recombination and has no core c-NHEJ phenotype•PAXX’s function in c-NHEJ is masked by XLF Tadi et al. find that PAXX interacts with Ku threaded on DNA via the Ku70 subunit and requires a bare DNA extension for stability. They show that PAXX’s function overlaps with c-NHEJ factor XLF, explaining why its ablation has only a mild effect on NHEJ. In mammalian cells, classical non-homologous end joining (c-NHEJ) is critical for DNA double-strand break repair induced by ionizing radiation and during V(D)J recombination in developing B and T lymphocytes. Recently, PAXX was identified as a c-NHEJ core component. We report here that PAXX-deficient cells exhibit a cellular phenotype uncharacteristic of a deficiency in c-NHEJ core components. PAXX-deficient cells display normal sensitivity to radiomimetic drugs, are proficient in transient V(D)J recombination assays, and do not shift toward higher micro-homology usage in plasmid repair assays. Although PAXX-deficient cells lack c-NHEJ phenotypes, PAXX forms a stable ternary complex with Ku bound to DNA. Formation of this complex involves an interaction with Ku70 and requires a bare DNA extension for stability. Moreover, the relatively weak Ku-dependent stimulation of LIG4/XRCC4 activity by PAXX is unmasked by XLF ablation. Thus, PAXX plays an accessory role during c-NHEJ that is largely overlapped by XLF's function.
Author	Roy, Sunetra Nemoz, Clément Drevet, Pascal Audebert, Stéphane Tellier-Lebègue, Carine Charbonnier, Jean-Baptiste Tadi, Satish K. Modesti, Mauro Meek, Katheryn
Author_xml	– sequence: 1 givenname: Satish K. surname: Tadi fullname: Tadi, Satish K. organization: Cancer Research Center of Marseille, CNRS UMR7258, INSERM U1068, Institut Paoli-Calmettes, Aix-Marseille Université UM105, 13273 Marseille, France – sequence: 2 givenname: Carine surname: Tellier-Lebègue fullname: Tellier-Lebègue, Carine organization: Institute for Integrative Biology of the Cell (I2BC), IBITECS, CEA, CNRS, University Paris-Sud, Université Paris-Saclay, 91198 Gif-sur-Yvette cedex, France – sequence: 3 givenname: Clément surname: Nemoz fullname: Nemoz, Clément organization: Institute for Integrative Biology of the Cell (I2BC), IBITECS, CEA, CNRS, University Paris-Sud, Université Paris-Saclay, 91198 Gif-sur-Yvette cedex, France – sequence: 4 givenname: Pascal surname: Drevet fullname: Drevet, Pascal organization: Institute for Integrative Biology of the Cell (I2BC), IBITECS, CEA, CNRS, University Paris-Sud, Université Paris-Saclay, 91198 Gif-sur-Yvette cedex, France – sequence: 5 givenname: Stéphane surname: Audebert fullname: Audebert, Stéphane organization: Cancer Research Center of Marseille, CNRS UMR7258, INSERM U1068, Institut Paoli-Calmettes, Aix-Marseille Université UM105, 13273 Marseille, France – sequence: 6 givenname: Sunetra surname: Roy fullname: Roy, Sunetra organization: Department of Microbiology & Molecular Genetics, and Department of Pathobiology & Diagnostic Investigation, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA – sequence: 7 givenname: Katheryn surname: Meek fullname: Meek, Katheryn organization: Department of Microbiology & Molecular Genetics, and Department of Pathobiology & Diagnostic Investigation, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824, USA – sequence: 8 givenname: Jean-Baptiste surname: Charbonnier fullname: Charbonnier, Jean-Baptiste organization: Institute for Integrative Biology of the Cell (I2BC), IBITECS, CEA, CNRS, University Paris-Sud, Université Paris-Saclay, 91198 Gif-sur-Yvette cedex, France – sequence: 9 givenname: Mauro surname: Modesti fullname: Modesti, Mauro email: mauro.modesti@inserm.fr organization: Cancer Research Center of Marseille, CNRS UMR7258, INSERM U1068, Institut Paoli-Calmettes, Aix-Marseille Université UM105, 13273 Marseille, France
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Keywords	Ku70 DNA double-strand break repair MMEJ XLF PAXX XRCC4 V(D)J recombination DNA ligase 4 Ku NHEJ
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Snippet	In mammalian cells, classical non-homologous end joining (c-NHEJ) is critical for DNA double-strand break repair induced by ionizing radiation and during V(D)J...
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SubjectTerms	B-Lymphocytes - metabolism DNA Breaks, Double-Stranded DNA double-strand break repair DNA End-Joining Repair - genetics DNA ligase 4 DNA Repair - genetics DNA Repair Enzymes - genetics DNA Repair Enzymes - metabolism DNA-Binding Proteins - chemistry DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism HCT116 Cells Humans Ku Autoantigen - chemistry Ku Autoantigen - genetics Ku Autoantigen - metabolism Ku70 Life Sciences MMEJ NHEJ PAXX T-Lymphocytes - metabolism V(D)J recombination V(D)J Recombination - genetics XLF XRCC4
Title	PAXX Is an Accessory c-NHEJ Factor that Associates with Ku70 and Has Overlapping Functions with XLF
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