title: DNA structure-specific priming of ATR activation by DNA-PKcs.
abstract: Three phosphatidylinositol-3-kinase-related protein kinases implement cellular responses to DNA damage. DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and ataxia-telangiectasia mutated respond primarily to DNA double-strand breaks (DSBs). Ataxia-telangiectasia and RAD3-related (ATR) signals the accumulation of replication protein A (RPA)-covered single-stranded DNA (ssDNA), which is caused by replication obstacles. Stalled replication intermediates can further degenerate and yield replication-associated DSBs. In this paper, we show that the juxtaposition of a double-stranded DNA end and a short ssDNA gap triggered robust activation of endogenous ATR and Chk1 in human cell-free extracts. This DNA damage signal depended on DNA-PKcs and ATR, which congregated onto gapped linear duplex DNA. DNA-PKcs primed ATR/Chk1 activation through DNA structure-specific phosphorylation of RPA32 and TopBP1. The synergistic activation of DNA-PKcs and ATR suggests that the two kinases combine to mount a prompt and specific response to replication-born DSBs.
authors.affiliation: Institute of Human Genetics, Unité Propre de Recherche 1142, Centre National de la Recherche Scientifique, 34396 Montpellier, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Vidal-Eychenié
authors.forename: Chantal
authors.initials: C
authors.lastname: Décaillet
authors.forename: Jihane
authors.initials: J
authors.lastname: Basbous
authors.forename: Angelos
authors.initials: A
authors.lastname: Constantinou
mesh_terms: D064007:Ataxia Telangiectasia Mutated Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D051747:DNA-Activated Protein Kinase / Q000378:metabolism*; D004789:Enzyme Activation; D006367:HeLa Cells; D006801:Humans; D009690:Nucleic Acid Conformation
chemical_list: D004247:DNA; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D051747:DNA-Activated Protein Kinase
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