Effects of DNA lesions on the transcription reaction of mitochondrial RNA polymerase: Implications for bypass RNA synthesis on oxidative DNA lesions

Nozomi Nakanishi, Atsushi Fukuoh, Dongchon Kang, Shigenori Iwai, Isao Kuraoka

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Oxidative DNA lesions inhibit the transcription of RNA polymerase II, but in the presence of transcription elongation factors, the transcription can bypass the lesions. Single-subunit mitochondrial RNA polymerase (mtRNAP) catalyses the synthesis of essential transcripts in mitochondria where reactive oxidative species (ROS) are generated as by-products. The occurrence of RNA synthesis by mtRNAP at oxidative DNA lesions remains unknown. Purified mtRNAP and a complex of RNA primer/DNA template containing a single DNA lesion, such as ROS-induced 8-oxoguanine (8-oxoG), two isomeric thymine glycols (5R-Tg or 5S-Tg), the UV-induced cis-syn cyclobutane pyrimidine dimer (CPD) and the pyrimidine(6-4)pyrimidone photoproduct (6-4pp), or a spontaneous common DNA lesion, a base-loss-induced apurinic/apyrimidinic (AP) site, were used for in vitro RNA synthesis assays. In this report, we show that mtRNAP bypassed the oxidative DNA lesions of non-bulky 8-oxoG and 5R-Tg and 5S-Tg with pausing sites but did not bypass the UV-induced DNA lesions and the AP site. The bacteriophage T7 phage RNA polymerase, which is homologous to mtRNAP, bypassed 8-oxoG but stalled at 5R-Tg and 5S-Tg. As expected, although translesion RNA synthesis in 8-oxoG on the DNA templates generated incorrect transcripts with a G:C to T:A transversion, the synthesis in Tg could lead to the correct transcripts with no transcriptional mutagenesis. Collectively, these data suggest that mtRNAP may tolerate the mitochondrial genome containing oxidative DNA lesions induced by ROS from the side effects of an ATP generation reaction.

Original languageEnglish
Pages (from-to)117-123
Number of pages7
JournalMutagenesis
Volume28
Issue number1
DOIs
Publication statusPublished - Jan 1 2013

Fingerprint

DNA-Directed RNA Polymerases
Transcription
RNA
DNA
Bacteriophages
Pyrimidinones
Bacteriophage T7
Pyrimidine Dimers
Peptide Elongation Factors
mitochondrial RNA
Cohort Effect
Mutagenesis
Mitochondria
Mitochondrial Genome
RNA Polymerase II
Byproducts
Assays
Transcription Factors
Genes
Adenosine Triphosphate

All Science Journal Classification (ASJC) codes

  • Genetics
  • Toxicology
  • Genetics(clinical)
  • Health, Toxicology and Mutagenesis

Cite this

Effects of DNA lesions on the transcription reaction of mitochondrial RNA polymerase : Implications for bypass RNA synthesis on oxidative DNA lesions. / Nakanishi, Nozomi; Fukuoh, Atsushi; Kang, Dongchon; Iwai, Shigenori; Kuraoka, Isao.

In: Mutagenesis, Vol. 28, No. 1, 01.01.2013, p. 117-123.

Research output: Contribution to journalArticle

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