In vivo determination of replication origins of human mitochondrial DNA by ligation-mediated polymerase chain reaction

Dongchon Kang, Kenichi Miyako, Yoichiro Kai, Takashi Irie, Koichiro Takeshige

Research output: Contribution to journalArticlepeer-review

79 Citations (Scopus)

Abstract

A large part of replication is aborted in human mitochondria, the result being a D-loop. As few attempts have been made to distinguish free 5' ends of true replicate from those of abortive ones, we examined the 5' ends of true replicate of human mitochondrial DNA at one nucleotide resolution in vivo by making use of ligation-mediated polymerase chain reaction. The distribution and relative amounts of origins of the true replicate are exactly the same as those of total newly synthesized heavy strands, which means that the abortion of replication is independent or 5' ends. Treatment of DNA with RNase H frees 5' ends on both heavy and light strands. This is the first in vivo evidence for covalently attached primer RNA to nascent strand in human mitochondrial DNA.

Original languageEnglish
Pages (from-to)15275-15279
Number of pages5
JournalJournal of Biological Chemistry
Volume272
Issue number24
DOIs
Publication statusPublished - Jun 13 1997

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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