Abstract
The muscle mitochondria of a patient with Kearns-Sayre/chronic external ophthalmoplegia plus syndrome were found to be completely deficient in respiratory complex I activity and partially deficient in complex IV and V activities. Treatment of the patient with coenzyme Q10 and succinate resulted in clinical improvement of respiratory function, consistent with the respiratory deficiencies. Restriction enzyme analysis of the muscle mtDNA revealed a 4.9-kilobase deletion in 50% of the mtDNA molecules. Polymerase chain reaction analysis demonstrated that the deletion was present in the patient's muscle but not in her lymphocytes or platelets. Furthermore, the deletion was not present in the muscle or platelets of two sisters. Hence, the mutation probably occurred in the patient's somatic cells. Direct sequencing of polymerase chain reaction-amplified DNA revealed a 4977-base-pair deletion removing four genes for subunits of complex I, one gene for complex IV, two genes for complex V, and five genes for tRNAs, which paralleled the respiratory enzymes affected in the disease. A 13-base-pair direct repeat was observed upstream from both breakpoints. Relative to the direction of heavy-strand replication, the first repeat was retained and the second repeat was deleted, suggesting a slip-replication mechanism. Sequence analysis of the human mtDNA revealed many direct repeats of 10 base pairs or greater, indicating that this mechanism could account for other reported deletions. We postulate that the prevalence of direct repeats in the mtDNA is a consequence of the guanine-cytosine bias of the heavy and light strands.
MeSH Terms
Base Sequence
Chromosome Deletion
DNA Replication
DNA, Mitochondrial/genetics
Electron Transport Complex IV/genetics
Female
Genes
Humans
Kearns-Sayre Syndrome/genetics,metabolism
Middle Aged
Mitochondria, Muscle/metabolism
Mitochondrial Proton-Translocating ATPases
Models, Genetic
Molecular Sequence Data
Muscles/metabolism,pathology
NAD(P)H Dehydrogenase (Quinone)
Ophthalmoplegia/genetics
Oxidative Phosphorylation
Polymerase Chain Reaction
Proton-Translocating ATPases/genetics
Quinone Reductases/genetics
Sequence Homology, Nucleic Acid
Chemicals
DNA, Mitochondrial
complex V (mitochondrial oxidative phosphorylation system)
NAD(P)H Dehydrogenase (Quinone)
Quinone Reductases
Electron Transport Complex IV
Mitochondrial Proton-Translocating ATPases
Proton-Translocating ATPases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Shoffner J M
Department of Neurology, Emory University School of Medicine, Atlanta, GA 30322.
Lott M T
Voljavec A S
Soueidan S A
Costigan D A
Wallace D C
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