Evidence for nonhomologous end joining and non allelic homologous recombination in atypical NF1 microdeletions

NF1 microdeletion syndrome is caused by haploinsufficiency of the NF1 gene and of gene(s) located in adjacent flanking regions. Most of the NF1 deletions originate by non-allelic homologous recombination between repeated sequences (REP-P and –M) mapped to 17q11.2, while the remaining show unusual breakpoints. We performed high-resolution FISH analysis of 18 NF1 microdeleted patients with the aims of mapping non-recurrent deletion breakpoints and verifying the presence of additional recombination-prone architectural motifs. This approach allowed us to obtain the sequence of the first junction fragment of an atypical deletion. By conventional FISH, we identified 16 patients with REP-mediated common deletions, and two patients carrying atypical deletions of 1.3 Mb and 3 Mb. Following fiber-FISH, we identified breakpoint regions of 100 Kb, which led to the generation of several locus-specific probes restricting the atypical deletion endpoint intervals to a few Kb. Sequence analysis evidenced small blocks of REPs, clustered around the 1.3 Mb deletion breakpoints, probably involved in intrachromatid non allelic homologous recombination (NAHR), while isolation and sequencing of the 3 Mb deletion junction fragment indicated that a non homologous end joining (NHEJ) mechanism is implicated.