DNA-mediated charge transport as a probe of MutY/DNA interaction

Elizabeth M. Boon, Mary Ann Pope, Scott D. Williams, Sheila S. David, Jacqueline K. Barton

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

MutY is an Escherichia coli DNA repair enzyme that binds to 8-oxo-G:A and G:A mismatches and catalyzes the deglycosylation of the mismatched 2′-deoxyadenosine. We have applied DNA-mediated charge transport to probe the interaction of MutY with its DNA substrate. Oligonucleotides synthesized with a tethered rhodium intercalator and guanine doublets placed before and after the MutY binding site are used to assay for base flipping activity by MutY. On the basis of this assay, we find no evidence that MutY uses progressive base flipping as a means to find its binding site; protein binding does not perturb long-range DNA charge transport. DNA-mediated charge transport can be utilized to promote protein-DNA cross-linking from a distance. Long-range oxidation of 8-oxo-G within the MutY binding site using tethered rhodium intercalators promoted cross-linking and yielded information on MutY side chains that interact with this base. On the basis of photooxidative cross-linking of the wild type but not K142A mutant, it is evident that, within the protein complex, lysine 142 makes important contacts with 8-oxo-G.

Original languageEnglish (US)
Pages (from-to)8464-8470
Number of pages7
JournalBiochemistry
Volume41
Issue number26
DOIs
StatePublished - Jul 2 2002
Externally publishedYes

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DNA Probes
Charge transfer
DNA
Intercalating Agents
Rhodium
Binding Sites
Assays
DNA Repair Enzymes
Guanine
Protein Binding
Oligonucleotides
Escherichia coli
Lysine
Proteins
Oxidation
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Boon, E. M., Pope, M. A., Williams, S. D., David, S. S., & Barton, J. K. (2002). DNA-mediated charge transport as a probe of MutY/DNA interaction. Biochemistry, 41(26), 8464-8470. https://doi.org/10.1021/bi012068c

DNA-mediated charge transport as a probe of MutY/DNA interaction. / Boon, Elizabeth M.; Pope, Mary Ann; Williams, Scott D.; David, Sheila S.; Barton, Jacqueline K.

In: Biochemistry, Vol. 41, No. 26, 02.07.2002, p. 8464-8470.

Research output: Contribution to journalArticle

Boon, EM, Pope, MA, Williams, SD, David, SS & Barton, JK 2002, 'DNA-mediated charge transport as a probe of MutY/DNA interaction', Biochemistry, vol. 41, no. 26, pp. 8464-8470. https://doi.org/10.1021/bi012068c
Boon EM, Pope MA, Williams SD, David SS, Barton JK. DNA-mediated charge transport as a probe of MutY/DNA interaction. Biochemistry. 2002 Jul 2;41(26):8464-8470. https://doi.org/10.1021/bi012068c
Boon, Elizabeth M. ; Pope, Mary Ann ; Williams, Scott D. ; David, Sheila S. ; Barton, Jacqueline K. / DNA-mediated charge transport as a probe of MutY/DNA interaction. In: Biochemistry. 2002 ; Vol. 41, No. 26. pp. 8464-8470.
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