The mtDNA mutation spectrum of the progeroid Polg mutator mouse includes abundant control region multimers.
Polg mtDNA mutator mice are important models for investigating the role of acquired mtDNA mutations in aging. Despite extensive study, there remains little consensus on either the etiology of the progeroid phenotype or the mtDNA mutation spectrum induced by disrupted polymerase-γ function. To investigate the latter, we have developed a novel, pragmatic approach we term "Mito-seq," applying next-generation sequencing to enriched, native mtDNA. Regardless of detection parameters we observed an increase of at least two orders of magnitude in the number of mtDNA single nucleotide variants in Polg mutator mice compared to controls. We found no evidence for the accumulation of canonical mtDNA deletions but multimers of the mtDNA control region were identified in brain and heart. These control region multimers (CRMs) contained heterogeneous breakpoints and formed species that excluded the majority of mtDNA genes. CRMs demonstrate that polymerase-γ 3'-5' exonuclease activity is required for preserving mtDNA integrity.
Department of Neurology, University of Miami Miller School of Medicine, Miami, FL 33136, USA.
Siôn L
SL
Williams
Jia
J
Huang
Yvonne J K
YJ
Edwards
Rick H
RH
Ulloa
Lloye M
LM
Dillon
Tomas A
TA
Prolla
Jeffery M
JM
Vance
Carlos T
CT
Moraes
Stephan
S
Züchner
D000375:Aging / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D000074002:DNA Polymerase gamma; D004272:DNA, Mitochondrial / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D019901:Locus Control Region / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D011371:Progeria / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*
D004272:DNA, Mitochondrial; D000074002:DNA Polymerase gamma; D004259:DNA-Directed DNA Polymerase; C498803:Polg protein, mouse