Human endonuclease III acts preferentially on DNA damage opposite guanine residues in DNA

L. Eide, L. Luna, E. C. Gustad, Paul Henderson, J. M. Essigmann, B. Demple, E. Seeberg

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

The human endonuclease III homologue (hNTH1) removes premutagenic cytosine damage from DNA. This includes 5-hydroxycytosine, which has increased potential for pairing with adenine, resulting in C → T transition mutations. Here we report that hNTH1 acts on both 5-hydroxycytosine and abasic sites preferentially when these are situated opposite guanines in DNA. Discrimination against other opposite bases is strongly dependent on the presence of magnesium. To further elucidate this effect, we have introduced mutations in the helix-hairpin-helix domain of hNTH1 (K212S, P211R, +G212, and ΔP211), and measured the kinetics of 5-hydroxycytosine removal of the mutants relative to wild type. The K212S and ΔP211 (truncated hairpin) mutant proteins were both inactive, whereas the extended hairpin in the +G212 mutant diminished recognition and binding to 5-hydroxycytosine-containing DNA. The P211R mutant resembled native hNTH1, except for decreased specificity of binding. Despite the altered kinetic parameters, the active mutants retained the ability to discriminate against the pairing base, indicating that enzyme interactions with the opposite strand relies on other domains than the active site helix-hairpin-helix motif.

Original languageEnglish (US)
Pages (from-to)6653-6659
Number of pages7
JournalBiochemistry
Volume40
Issue number22
DOIs
StatePublished - Jun 5 2001
Externally publishedYes

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Endonucleases
Guanine
DNA Damage
DNA
Mutation
Cytosine
Adenine
Mutant Proteins
Kinetic parameters
Base Pairing
Magnesium
Catalytic Domain
Kinetics
5-hydroxycytosine
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Eide, L., Luna, L., Gustad, E. C., Henderson, P., Essigmann, J. M., Demple, B., & Seeberg, E. (2001). Human endonuclease III acts preferentially on DNA damage opposite guanine residues in DNA. Biochemistry, 40(22), 6653-6659. https://doi.org/10.1021/bi0028901

Human endonuclease III acts preferentially on DNA damage opposite guanine residues in DNA. / Eide, L.; Luna, L.; Gustad, E. C.; Henderson, Paul; Essigmann, J. M.; Demple, B.; Seeberg, E.

In: Biochemistry, Vol. 40, No. 22, 05.06.2001, p. 6653-6659.

Research output: Contribution to journalArticle

Eide, L, Luna, L, Gustad, EC, Henderson, P, Essigmann, JM, Demple, B & Seeberg, E 2001, 'Human endonuclease III acts preferentially on DNA damage opposite guanine residues in DNA', Biochemistry, vol. 40, no. 22, pp. 6653-6659. https://doi.org/10.1021/bi0028901
Eide, L. ; Luna, L. ; Gustad, E. C. ; Henderson, Paul ; Essigmann, J. M. ; Demple, B. ; Seeberg, E. / Human endonuclease III acts preferentially on DNA damage opposite guanine residues in DNA. In: Biochemistry. 2001 ; Vol. 40, No. 22. pp. 6653-6659.
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