Summary The Mre11/Rad50/NBS1 (MRN) complex plays many roles in response to DNA double strand breaks (DSBs), but its functions in repair by non homologous end joining (NHEJ) pathways are poorly understood. We have investigated requirements for MRN in Class Switch Recombination (CSR), a programmed DNA rearrangement in B lymphocytes that requires NHEJ. To this end we have engineered mice that lack the entire MRN complex in B lymphocytes, or possess an intact complex harboring mutant Mre11 lacking DNA nuclease activities. MRN deficiency confers a striking defect in CSR, impacting both the Classic and Alternative NHEJ pathways. In contrast, absence of Mre11 nuclease activities causes a milder phenotype, revealing a separation of function within the complex. We propose a model in which MRN stabilizes distant breaks and processes DNA termini to facilitate repair by both the Classical and Alternative NHEJ pathways.

Homologous recombination (HR) facilitates accurate repair of DNA double strand breaks (DSBs) during S and G2 phases of the cell cycle by using intact sister chromatids as sequence templates. HR capacity is maximized in S and G2 by Cyclin–Dependent Kinase (CDK) phosphorylation of CtIP, which subsequently interacts with BRCA1 and the Mre11–Rad50–NBS1 (MRN) complex. Here we show that Mre11 controls these events through a direct interaction with CDK2 that is required for CtIP phosphorylation and BRCA1 interaction in normally dividing cells. CDK2 binds the C–terminus of Mre11, which is absent in an inherited allele causing Ataxia–Telangiectasia Like Disorder. This newly uncovered role for Mre11 does not require ATM activation or nuclease activities. Therefore, functions of MRN are not restricted to DNA damage responses, but include regulating HR capacity during the normal mammalian cell cycle.

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