Mitochondrial DNA replication proceeds via a 'bootlace' mechanism involving the incorporation of processed transcripts.

TitleMitochondrial DNA replication proceeds via a 'bootlace' mechanism involving the incorporation of processed transcripts.
Publication TypeJournal Article
Year of Publication2013
AuthorsReyes, A, Kazak, L, Wood, SR, Yasukawa, T, Jacobs, HT, Holt, IJ
JournalNucleic Acids Res
Volume41
Issue11
Pagination5837-50
Date Published2013 Jun
ISSN1362-4962
KeywordsAnimals, Cross-Linking Reagents, Deoxyadenine Nucleotides, Deoxyribonucleotides, DNA Replication, DNA, Mitochondrial, Ficusin, Humans, Mice, Mice, Inbred BALB C, RNA, RNA Precursors, RNA, Mitochondrial
Abstract

The observation that long tracts of RNA are associated with replicating molecules of mitochondrial DNA (mtDNA) suggests that the mitochondrial genome of mammals is copied by an unorthodox mechanism. Here we show that these RNA-containing species are present in living cells and tissue, based on interstrand cross-linking. Using DNA synthesis in organello, we demonstrate that isolated mitochondria incorporate radiolabeled RNA precursors, as well as DNA precursors, into replicating DNA molecules. RNA-containing replication intermediates are chased into mature mtDNA, to which they are thus in precursor-product relationship. While a DNA chain terminator rapidly blocks the labeling of mitochondrial replication intermediates, an RNA chain terminator does not. Furthermore, processed L-strand transcripts can be recovered from gel-extracted mtDNA replication intermediates. Therefore, instead of concurrent DNA and RNA synthesis, respectively, on the leading and lagging strands, preformed processed RNA is incorporated as a provisional lagging strand during mtDNA replication. These findings indicate that RITOLS is a physiological mechanism of mtDNA replication, and that it involves a 'bootlace' mechanism, in which processed transcripts are successively hybridized to the lagging-strand template, as the replication fork advances.

DOI10.1093/nar/gkt196
Alternate JournalNucleic Acids Res.
Citation Key10.1093/nar/gkt196
PubMed ID23595151
PubMed Central IDPMC3675460
Grant ListMC_U105663140 / / Medical Research Council / United Kingdom