Impact of the m.13513G>A Variant on the Functions of the OXPHOS System and Cell Retrograde Signaling

Mitochondria are involved in many vital functions in living cells, including the synthesis of ATP by oxidative phosphorylation (OXPHOS) and regulation of nuclear gene expression through retrograde signaling. Leigh syndrome is a heterogeneous neurological disorder resulting from an isolated complex I deficiency that causes damage to mitochondrial energy production. The pathogenic mitochondrial DNA (mtDNA) variant m.13513G>A has been associated with Leigh syndrome. The present study investigated the effects of this mtDNA variant on the OXPHOS system and cell retrograde signaling. Transmitochondrial cytoplasmic hybrid (cybrid) cell lines harboring 50% and 70% of the m.13513G>A variant were generated and tested along with wild-type (WT) cells. The functionality of the OXPHOS system was evaluated by spectrophotometric assessment of enzyme activity and high-resolution respirometry. Nuclear gene expression was investigated by RNA sequencing and droplet digital PCR. Increasing levels of heteroplasmy were associated with reduced OXPHOS system complex I, IV, and I + III activities, and high-resolution respirometry also showed a complex I defect. Profound changes in transcription levels of nuclear genes were observed in the cell lines harboring the pathogenic mtDNA variant, indicating the physiological processes associated with defective mitochondria.

Automated summary

Mitochondria play essential roles in diverse cellular functions, such as ATP synthesis and regulation of nuclear gene expression. Leigh syndrome is a neurological disorder caused by complex I deficiency, leading to impaired mitochondrial energy production. The mtDNA variant m.13513G>A has been associated with Leigh syndrome. This study examined the impact of this mtDNA variant on the OXPHOS system and cell retrograde signaling. The results showed that increasing levels of the variant were associated with reduced OXPHOS system activity and complex I defect, as well as profound changes in nuclear gene expression.