Mutagenic effects of 5-formyluracil on a plasmid vector during replication in Escherichia coli.

Abstract:

PURPOSE:5-Formyluracil (5-foU) is a major derivative of thymine produced in DNA by ionizing radiation and various chemical oxidants. It has been previously shown that 5-foU in template DNA directs misincorporation of nucleotides by DNA polymerases during in vitro DNA synthesis. The present experiments were designed to understand the biological effects of5-foU in vivo. MATERIALS AND METHODS:The modified base was incorporated site-specifically into the recognition site of restriction endonuclease SalI (5'-GTCGAC) or AflII (5'-CTTAAG) in vector plasmid pSVK3 and introduced the plasmid into Escherichia coli. RESULTS:When the plasmids were replicated in E. coli, 5-foU caused mutations at the target sites. The induced mutation frequencies were 0.038-0.049%. Sequence analysis revealed that 5-foU preferentially caused T:A-->C:G and T:A-->A:T base substitutions and -1 deletions at the 5-foU site. 5-FoU also caused mutations at sites near the 5-foU. The alkA mutation did not affect the frequency of mutations in 5-foU-containing plasmids. CONCLUSIONS:The present experiments demonstrated that 5-formyluracil in DNA caused mutations in E. coli.

journal_name

Int J Radiat Biol

authors

Miyabe I,Zhang QM,Sugiyama H,Kino K,Yonei S

doi

10.1080/095530001453113

subject

Has Abstract

pub_date

2001-01-01 00:00:00

pages

53-8

issue

1

eissn

0955-3002

issn

1362-3095

journal_volume

77

pub_type

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