Cleavage of the BRCT tandem domains of nibrin by the 657del5 mutation affects the DNA damage response less than the Arg215Trp mutation

The Nijmegen breakage syndrome (NBS) is a genetic disorder. caused by mutations in NBN gene and characterized by chromosomal. instability and hypersensitivity to ionizing radiations (IR).. The N-terminus of nibrin (NBN) contains a tandem breast cancer. 1 (BRCA1) carboxy-terminal (BRCT) domain that represents one. of the major mediators of phosphorylation-dependent protein-protein. interactions in processes related to cell cycle checkpoint and. DNA repair functions. Patients with NBS compound heterozygous. for the 657del5 hypomorphic mutation and for the Arg215Trp. missense mutation (corresponding to the 643C[T gene mutation). display a clinical phenotype more severe than that of patients. homozygous for the 657del5 mutation. Here, we show that both. the 657del5 and Arg215Trp mutations, occurring within the tandem. BRCT domains of NBN, although not altering the assembly. of the MRE11\\\/RAD50\\\/NBN (MRN) complex, affect the MRE11. IR-induced nuclear foci (IRIF) formation and the DNA doublestrand. break (DSB) signaling via the phosphorylation of both. ataxia-telangiectasia-mutated (ATM) kinase and ATM downstream. targets (e.g., SMC1 and p53). Remarkably, data obtained. indicate that the cleavage of the BRCT tandem domains of NBN. by the 657del5 mutation affects the DNA damage response less. than the Arg215Trp mutation. Indeed, the 70-kDa NBN fragment,. arising from the 657del5 mutation, maintains the capability. to interact with MRE11 and c-H2AX and to form IRIF. Altogether,. the role of the tandem BRCT domains of NBN in the. localization of the MRN complex at the DNA DSB and in the. activation of the damage response is highlighted.