Figure 6. The human MARS2 mutations.

(A) PCR amplification products of MARS2 encompassing a portion of the coding sequence revealed the presence of a 268 bp deletion mutation segregating in ARSAL Family E but not in Family B. This truncated product is indicated by an arrow. The normal PCR product is around 500 bp. Segregation of the deletion is shown in Family E; brothers E10 and E11 carry the mutation. Their unaffected father E9 is also a carrier. The determined genotypes for the patients shown (summarized in Table S5 for all patients) are shown above the PCR bands. (B) Wild-type sequence of MARS2 PCR products. (C) DNA sequencing of the deletion (c.681Δ268bpfx236X). (D–E) Nonrecurrent rearrangements involving the MARS2 gene was confirmed by the oligonucleotide custom aCGH. In patients E10 and E11, the array discriminated the presence of the duplication as well as the deletion (see arrows) as depicted by the lower band detecting only one additional copy. (F) PCR amplification products of MARS2 encompassing the coding sequence revealed the presence of a ∼300 bp insertion mutation segregating in ARSAL family members C6 and C8 but not in Family B. This larger amplification product is indicated by an arrow. The normal amplicon size is about 800 bp. C5 is the unaffected father of C6 and C8 and also carries the mutation. (G) Wild-type sequence of MARS2. (H) DNA sequencing of the heterozygous case C6 corresponding to the insertion revealed parts of the MARS2 duplication mutation. Rearrangement was confirmed by oligonucleotide custom aCGH. Note that the array data of C6, a compound heterozygote (Dup2/Dup2), demonstrates the presence of a potentially larger duplication while not showing the 300 bp insertion, the array not having been designed to include its sequence. (I) In homozygous patient B4 (Dup1/Dup1), the array suggests that the duplication has identical distal and proximal breakpoint junctions with the other ARSAL cases.