title: 53BP1 and NFBD1/MDC1-Nbs1 function in parallel interacting pathways activating ataxia-telangiectasia mutated (ATM) in response to DNA damage.
abstract: 53BP1 and NFBD1/MDC1 are recruited rapidly to sites of DNA double-strand breaks (DSBs), where they are hypothesized to function downstream of the ataxia-telangiectasia mutated (ATM) checkpoint kinase as "mediators" of DNA DSB signaling. To test this hypothesis, we suppressed 53BP1 and NFBD1/MDC1 expression by small interference RNA and monitored ATM autophosphorylation at Ser(1981) as a marker for ATM activation. Suppression of NFBD1/MDC1 led to decreased ATM activation and phosphorylation of ATM substrates. This phenotype was identical to that observed in cells with defective Nbs1 function and is consistent with recent observations identifying NFBD1/MDC1 as a component of the Mre11-Rad50-Nbs1 protein complex. In cells with wild-type Nbs1, suppression of 53BP1 expression had no effect on ATM activation but was associated with increased recruitment of NFBD1/MDC1 and Nbs1 to sites of DNA breaks, suggesting that decreased 53BP1 function might be compensated for by increased NFBD1/MDC1 and Nbs1 activity. Indeed, in cells with mutant Nbs1, suppression of 53BP1 led to decreased ATM activation and phosphorylation of ATM substrates. We conclude that DNA DSBs activate ATM through at least two independent pathways involving 53BP1 and NFBD1/MDC1-Nbs1, respectively.
authors.affiliation: The Wistar Institute, Philadelphia, Pennsylvania 19104-4268, USA.
authors.forename: Tamara A
authors.initials: TA
authors.lastname: Mochan
authors.forename: Monica
authors.initials: M
authors.lastname: Venere
authors.forename: Richard A
authors.initials: RA
authors.lastname: DiTullio
authors.forename: Thanos D
authors.initials: TD
authors.lastname: Halazonetis
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D064007:Ataxia Telangiectasia Mutated Proteins; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000502:physiology*; D002460:Cell Line; D045744:Cell Line, Tumor; D064447:Checkpoint Kinase 2; D004249:DNA Damage / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D004789:Enzyme Activation; D005347:Fibroblasts; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D010750:Phosphoproteins; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D015534:Trans-Activators / Q000037:antagonists & inhibitors / Q000502:physiology*; D014162:Transfection; D025521:Tumor Suppressor Proteins; D000071857:Tumor Suppressor p53-Binding Protein 1
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; C413163:MDC1 protein, human; C112156:NBN protein, human; D009687:Nuclear Proteins; D010750:Phosphoproteins; D034741:RNA, Small Interfering; C498301:TP53BP1 protein, human; D015534:Trans-Activators; D025521:Tumor Suppressor Proteins; D000071857:Tumor Suppressor p53-Binding Protein 1; D064447:Checkpoint Kinase 2; C576324:ATM protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; C578634:CHEK2 protein, human; D017346:Protein Serine-Threonine Kinases