NDUFS6 mutations are a novel cause of lethal neonatal mitochondrial complex I deficiency
J. Clin. Invest. Denise M. Kirby, et al. 114:837 doi:10.1172/JCI20683 [
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Figure 5Analysis of
NDUFS6 cDNA and genomic DNA in families B and C. (
A) Two percent agarose gel of
NDUFS6 cDNA fragments generated by standard PCR. Lanes show control (Co); patient B (B); patient C (C); patient H (H); dH
2O blank (Bl). PCR of
NDUFS6 cDNA gave no product in patient B and a larger fragment than the control in patient C. (
B) A representation depicting the frameshift and creation of a premature stop codon caused by the 26-bp insertion of intronic sequence in the proband from family C (bottom). Translation of the control sequence is also shown for comparison (top). The diagram shows the exon 2/exon 3 boundary, with the inserted sequence shown in b and underlined and a stop codon represented by an asterisk. (
C) 0.8% agarose gel of
NDUFS6 fragments generated by long-range PCR using forward and reverse primers located 6.2 kb apart. Lanes 2_5, unrelated complex I_deficient patients. The PCR product in patient B is approximately 4 kb smaller than that of other samples. The approximately 2-kb fragment from lane 6 was sequenced and the deletion breakpoints elucidated. (
D) Schematic representation of the 4.175-kb deletion, which encompasses exons 3 and 4 (shaded box) of the 14.5-kb
NDUFS6 gene in the proband from family B. The exact homozygous deletion breakpoints are also shown with the immediate sequence before and after each breakpoint included.
