Mitochondrial DNA depletion syndrome due to mutations in the RRM2B gene

Abstract

Mitochondrial DNA depletion syndrome (MDS) is characterized by a reduction in mtDNA copy number and has been associated with mutations in eight nuclear genes, including enzymes involved in mitochondrial nucleotide metabolism (POLG, TK2, DGUOK, SUCLA2, SUCLG1, PEO1) and MPV17. Recently, mutations in the RRM2B gene, encoding the p53-controlled ribonucleotide reductase subunit, have been described in seven infants from four families, who presented with various combinations of hypotonia, tubulopathy, seizures, respiratory distress, diarrhea, and lactic acidosis. All children died before 4 months of age. We sequenced the RRM2B gene in three unrelated cases with unexplained severe mtDNA depletion. The first patient developed intractable diarrhea, profound weakness, respiratory distress, and died at 3 months. The other two unrelated patients had a much milder phenotype and are still alive at ages 27 and 36 months. All three patients had lactic acidosis and severe depletion of mtDNA in muscle. Muscle histochemistry showed RRF and COX deficiency. Sequencing the RRM2B gene revealed three missense mutations and two single nucleotide deletions in exons 6, 8, and 9, confirming that RRM2B mutations are important causes of MDS and that the clinical phenotype is heterogeneous and not invariably fatal in infancy.

Figure 1

Modified Gomori trichrome stain of a muscle specimen (cross section) from patient 1 shows many ragged-red fibers (arrows). There are also numerous atrophic fibers and increased perimysial and endomysial connective tissue. X120.

Figure 2

The 3D structure of the wild-type protein (left upper panel) shows in red the part of this subunit missing in the mutant protein (right upper panel) due to the nonsense L208X mutation in patient 3. A detail of the wild-type conformation (left lower panel) shows how the spatial proximity of residue Ile224 (highlighted in yellow) to the Tyr138 residue (highlighted in red) could explain the effect of the I224S mutation (right lower panel) on the free radical position in the active pocket.

Figure 3

Alignment of the R2 subunit in human and three other species showing the positions of the α-helices (denoted as A-H) and the three missense mutations found in our patients (wild-type amino acids are highlighted and circled in red). Note that all mutations affect evolutionarily highly conserved sites.