Distinct Repair Activities of Human 7,8-Dihydro-8-oxoguanine DNA Glycosylase and Formamidopyrimidine DNA Glycosylase for Formamidopyrimidine and 7,8-Dihydro-8-oxoguanine*

7,8-Dihydro-8-oxoguanine (8-oxoG) and 2,6-diamino-4-hydroxyformamidopyrimidine (Fapy) are major DNA lesions formed by reactive oxygen species and are involved in mutagenic and/or lethal events in cells. Both lesions are repaired by human 7,8-dihydro-8-oxoguanine DNA glycosylase (hOGG1) and formamidopyrimidine DNA glycosylase (Fpg) in human andEscherichia coli cells, respectively. In the present study, the repair activities of hOGG1 and Fpg were compared using defined oligonucleotides containing 8-oxoG and a methylated analog of Fapy (me-Fapy) at the same site. Thek cat/K m values of hOGG1 for 8-oxoG and me-Fapy were comparable, and this was also the case for Fpg. However, the k cat/K m values of hOGG1 for both lesions were approximately 80-fold lower than those of Fpg. Analysis of the Schiff base intermediate by NaBH4trapping implied that lower substrate affinity and slower hydrolysis of the intermediate for hOGG1 than Fpg accounted for the difference. hOGG1 and Fpg showed distinct preferences of the base opposite 8-oxoG, with the activity differences being 19.8- (hOGG1) and 12-fold (Fpg) between the most and least preferred bases. Surprisingly, such preferences were almost abolished and less than 2-fold for both enzymes when me-Fapy was a substrate, suggesting that, unlike 8-oxoG, me-Fapy is not subjected to paired base-dependent repair. The repair efficiency of me-Fapy randomly incorporated in M13 DNA varied at the sequence level, but orders of preferred and unpreferred repair sites were quite different for hOGG1 and Fpg. The distinctive activities of hOGG1 and Fpg including enzymatic parameters (k cat/K m ), paired base, and sequence context effects may originate from the differences in the inherent architecture of the DNA binding domain and catalytic mechanism of the enzymes.

• Publication year

  2000

• Venue

  Journal of Biological Chemistry

• Publication date

  2000-02-18

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1074/JBC.275.7.4956PMID 10671534
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• Repair of me-Fapy in M13 DNA varies by sequence context, and the favored and disfavored repair sites differ between hOGG1 and Fpg.

  Confidence 0.93

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• Base preference opposite 8-oxoG is strong for both enzymes, but these preferences are largely lost for me-Fapy.

  Confidence 0.97

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• The catalytic efficiency of hOGG1 for both 8-oxoG and me-Fapy is about 80-fold lower than that of Fpg.

  Confidence 0.95

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• hOGG1 shows comparable catalytic efficiency for 8-oxoG and me-Fapy, and Fpg shows the same pattern for the two lesions.

  Confidence 0.96

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review

• 8-oxog

  DNA lesion

  7,8-dihydro-8-oxoguanine, an oxidized guanine DNA lesion examined as a substrate in the repair assays.

  Aliases: 7,8-dihydro-8-oxoguanine, 8-oxoguanine

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• fpg

  enzyme

  Formamidopyrimidine DNA glycosylase, a DNA repair enzyme used here as the bacterial counterpart for lesion excision.

  Aliases: formamidopyrimidine DNA glycosylase

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• hogg1

  enzyme

  Human 7,8-dihydro-8-oxoguanine DNA glycosylase, an enzyme that excises oxidized guanine lesions from DNA.

  Aliases: human 7,8-dihydro-8-oxoguanine DNA glycosylase

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• kcat/km

  kinetic parameter

  The catalytic efficiency metric used to compare enzyme activity on the defined DNA substrates.

  Aliases: catalytic efficiency

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• m13 dna

  DNA substrate

  A single-stranded phage DNA substrate used to evaluate lesion repair across different sequence sites.

  Aliases: M13 phage DNA

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• me-fapy

  DNA lesion

  A methylated analog of formamidopyrimidine used as a defined DNA lesion substrate in the repair assays.

  Aliases: methylated Fapy, methylated analog of Fapy

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• paired base preference

  sequence effect

  The dependence of repair activity on which DNA base is opposite the lesion site.

  Aliases: opposite base preference, paired-base dependence

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review
• sequence context

  sequence effect

  The local DNA sequence surrounding a lesion site that can influence repair efficiency.

  Aliases: local sequence context

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) review

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