Characterization of the test sample set.

<p>A) Samples from previously reported patients with Alu insertion in <i>MAK</i> exon 9 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142614#pone.0142614.ref021" target="_blank">21</a>] and control samples were PCR amplified to detect homozygous alleles for Alu insertion and WT alleles. B) Sequence of the inserted element (280 bp Alu, 54 bp poly-A and 13 bp duplication of exon 9 sequence). C) Sanger sequence of the exon 9 Alu insertion breakpoints.</p

PCR validation of Alu insertion identified by <i>in silico</i> analysis in patients from the discovery cohort.

<p>A) PCR amplification using primers spanning exon 9 (top) and nested PCR using Alu-specific primer (bottom). The 1,194 bp amplicon containing the Alu insertion (arrow) is present strongly in the homozygous sample and weakly in the heterozygous samples (top); the Alu insertion-specific amplification (491 bp, bottom) confirms the presence of the Alu insertion. B) Pedigree of patient D379_148, carrying a missense mutation (p.Gly16Arg) and the Alu insertion mutation. C) Evolutionary conservation of glycine 16, mutated in the patient D379_148. D) Protein domains in MAK and location of the p.Gly16Arg change. The mutation annotations are based on the NM_001242957 transcript, where A from the ATG start codon is designated as a +1 position.</p

Identification of Homozygous and Heterozygous <i>MAK</i>-Alu Insertions in a Discovery Sample Set.

<p>Identification of Homozygous and Heterozygous <i>MAK</i>-Alu Insertions in a Discovery Sample Set.</p

Alignment of standard BWA-based Illumina reads of a control (top) and a <i>MAK</i>-Alu homozygous (bottom) sample.

<p><i>MAK</i>-Alu alignment produces a coverage gap in exon 9 but does not clearly identify an insertion.</p

Specificity and Sensitivity of <i>In Silico</i> Method to Detect the <i>MAK</i>-Alu insertion.

<p>Specificity and Sensitivity of <i>In Silico</i> Method to Detect the <i>MAK</i>-Alu insertion.</p

A novel <i>HSD17B10</i> mutation impairing the activities of the mitochondrial RNase P complex causes X-linked intractable epilepsy and neurodevelopmental regression

<p>We report a Caucasian boy with intractable epilepsy and global developmental delay. Whole-exome sequencing identified the likely genetic etiology as a novel p.K212E mutation in the X-linked gene <i>HSD17B10</i> for mitochondrial short-chain dehydrogenase/reductase SDR5C1. Mutations in <i>HSD17B10</i> cause the HSD10 disease, traditionally classified as a metabolic disorder due to the role of SDR5C1 in fatty and amino acid metabolism. However, SDR5C1 is also an essential subunit of human mitochondrial RNase P, the enzyme responsible for 5′-processing and methylation of purine-9 of mitochondrial tRNAs. Here we show that the p.K212E mutation impairs the SDR5C1-dependent mitochondrial RNase P activities, and suggest that the pathogenicity of p.K212E is due to a general mitochondrial dysfunction caused by reduction in SDR5C1-dependent maturation of mitochondrial tRNAs.</p