Replication bypass and mutagenic effect of α-deoxyadenosine site-specifically incorporated into single-stranded vectors

Hironori Shimizu, Ryohei Yagi, Yoshiharu Kimura, Keisuke Makino, Hiroaki Terato, Yoshihiko Ohyama, Hiroshi Ide

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

α-2'-Deoxyadenosine (α) is a major adenine lesion produced by γ-ray irradiation of DNA under anoxic conditions. In this study, single-stranded recombinant M13 vectors containing α were constructed and transfected into Escherichia coli to assess lethal and mutagenic effects of this lesion. The data for α were further compared with those obtained with M13 vectors containing normal A or a model abasic site (F) at the same site. The transfection assay revealed that α constituted a moderate block to DNA replication. The in vivo replication capacity to pass through α was ~ 20% relative to normal A, but 20-fold higher than that of F constituting an almost absolute replication block. Similar data were obtained by in vitro replication of oligonucleotide templates containing α or F by E. coli DNA polymerase I. The mutagenic consequence of replicating M13 DNA containing α was analyzed by direct DNA sequencing of progeny phage. Mutagenesis was totally targeted at the site of α introduced into the vector. Mutation was exclusively a single nucleotide deletion and no base substitutions were detected. The deletion frequency associated α was dependent on the 3'-nearest neighbor base: with the 3'-nearest neighbor base T mutation (deletion) frequency was 26%, whereas 1% with the 3'-nearest neighbor base G. A possible mechanism of the single nucleotide deletion associated with α is discussed on the basis of the misinsertion-strand slippage model.

Original languageEnglish
Pages (from-to)597-603
Number of pages7
JournalNucleic Acids Research
Volume25
Issue number3
DOIs
Publication statusPublished - Jul 30 1997
Externally publishedYes

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Nucleotides
Escherichia coli
DNA Polymerase I
DNA
Adenine
Mutation Rate
DNA Replication
DNA Sequence Analysis
Oligonucleotides
Mutagenesis
Bacteriophages
Transfection
Mutation
2'-deoxyadenosine
In Vitro Techniques

ASJC Scopus subject areas

  • Genetics

Cite this

Replication bypass and mutagenic effect of α-deoxyadenosine site-specifically incorporated into single-stranded vectors. / Shimizu, Hironori; Yagi, Ryohei; Kimura, Yoshiharu; Makino, Keisuke; Terato, Hiroaki; Ohyama, Yoshihiko; Ide, Hiroshi.

In: Nucleic Acids Research, Vol. 25, No. 3, 30.07.1997, p. 597-603.

Research output: Contribution to journalArticle

Shimizu, Hironori ; Yagi, Ryohei ; Kimura, Yoshiharu ; Makino, Keisuke ; Terato, Hiroaki ; Ohyama, Yoshihiko ; Ide, Hiroshi. / Replication bypass and mutagenic effect of α-deoxyadenosine site-specifically incorporated into single-stranded vectors. In: Nucleic Acids Research. 1997 ; Vol. 25, No. 3. pp. 597-603.
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