KsgA, a 16S rRNA adenine methyltransferase, has a novel DNA glycosylase/ AP lyase activity to prevent mutations in Escherichia coli

Qiu Mei Zhang-Akiyama, Hironobu Morinaga, Masahiro Kikuchi, Shinichirou Yonekura, Hiroshi Sugiyama, Kazuo Yamamoto, Shuji Yonei

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The 5-formyluracil (5-foU), a major mutagenic oxidative damage of thymine, is removed from DNA by Nth, Nei and MutM in Escherichia coli. However, DNA polymerases can also replicate past the 5-foU by incorporating C and G opposite the lesion, although the mechanism of correction of the incorporated bases is still unknown. In this study, using a borohydride-trapping assay, we identified a protein trapped by a 5-foU/C-containing oligonucleotide in an extract from E. coli mutM nth nei mutant. The protein was subsequently purified from the E. coli mutM nth nei mutant and was identified as KsgA, a 16S rRNA adenine methyltransferase. Recombinant KsgA also formed the trapped complex with 5-foU/C- and thymine glycol (Tg)/C-containing oligonucleotides. Furthermore, KsgA excised C opposite 5-foU, Tg and 5-hydroxymethyluracil (5-hmU) from duplex oligonucleotides via a β-elimination reaction, whereas it could not remove the damaged base. In contrast, KsgA did not remove C opposite normal bases, 7,8-dihydro-8-oxoguanine and 2-hydroxyadenine. Finally, the introduction of the ksgA mutation increased spontaneous mutations in E. coli mutM mutY and nth nei mutants. These results demonstrate that KsgA has a novel DNA glycosylase/ AP lyase activity for C mispaired with oxidized T that prevents the formation of mutations, which is in addition to its known rRNA adenine methyltransferase activity essential for ribosome biogenesis.

Original languageEnglish
Pages (from-to)2116-2125
Number of pages10
JournalNucleic Acids Research
Volume37
Issue number7
DOIs
Publication statusPublished - 2009
Externally publishedYes

Fingerprint

DNA-(Apurinic or Apyrimidinic Site) Lyase
DNA Glycosylases
Methyltransferases
Adenine
Escherichia coli
Mutation
Oligonucleotides
Borohydrides
Thymine
DNA-Directed DNA Polymerase
Ribosomes
Proteins
5-formyluracil
DNA

ASJC Scopus subject areas

  • Genetics

Cite this

KsgA, a 16S rRNA adenine methyltransferase, has a novel DNA glycosylase/ AP lyase activity to prevent mutations in Escherichia coli. / Zhang-Akiyama, Qiu Mei; Morinaga, Hironobu; Kikuchi, Masahiro; Yonekura, Shinichirou; Sugiyama, Hiroshi; Yamamoto, Kazuo; Yonei, Shuji.

In: Nucleic Acids Research, Vol. 37, No. 7, 2009, p. 2116-2125.

Research output: Contribution to journalArticle

Zhang-Akiyama, Qiu Mei ; Morinaga, Hironobu ; Kikuchi, Masahiro ; Yonekura, Shinichirou ; Sugiyama, Hiroshi ; Yamamoto, Kazuo ; Yonei, Shuji. / KsgA, a 16S rRNA adenine methyltransferase, has a novel DNA glycosylase/ AP lyase activity to prevent mutations in Escherichia coli. In: Nucleic Acids Research. 2009 ; Vol. 37, No. 7. pp. 2116-2125.
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AU - Kikuchi, Masahiro

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