A) BAB2543 carries two duplications in an inverted
orientation separated with a copy-neutral segment (DUP-NML-DUP/INV). Breakpoint
junction (jct) 1 maps to inverted SMSREP LCRs, and evaded sequencing attempts.
Breakpoint jct 2 was mediated by inverted Alu repeats and forms
an Alu-Alu chimera; junction sequence is characterized by 29 bp
of microhomology. Eight de novo mutations have also been characterized within
17p11.2; Sanger sequencing electropherogram confirming each SNV is shown along
with the location, genomic context and type. B) BAB1931 carries
three deletions interspersed with copy-neutral segments (DEL-NML-DEL-NML-DEL).
SV breakpoints display one, two and zero bp of microhomology at the junctions,
and jct3 was previously uncharacterized. The four de novo SNVs and indels
present in the proband and Sanger sequencing electropherogram confirmation are
depicted below. The SNV at 20409881 was not independently confirmed by using a
PCR/Sanger sequencing strategy due to its presence within an LCR; however, it
was observed in both Illumina and PacBio sequencing data and shown to be de novo
in the trio Illumina sequencing data. C) Plot shows the relative
contribution of each SNV transition and transversion observed de novo in the
non-recurrent individuals. Overall abundance of C>G mutations can be
readily observed. D) Enrichment of de novo SNVs in proximity to SV
breakpoints was observed in the genomes of 9 out of 13 subjects with
non-recurrent SV. This enrichment was not observed for de novo SNVs (N=4)
detected in the subjects carrying recurrent SVs. The normalized statistics
(Z-value) for each simulation and observation (red dot) is displayed with the
box plots. E) Mutational clustering was examined in individuals
with more than one de novo SNV. SNV mutations show statically significant
clustering in 5 out of 9 NR rearrangements. The normalized statistics (Z-value)
for each simulation and the observation (red dot) are plotted. The box plots
were colored according to the number of de novo mutations detected in each
subject. (*) P ≤ 0.05; (**) P ≤ 0.01; (***) P ≤ 0.001. See
also STAR Methods, Figure S2 and Data S1.