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DNA-Mediated Charge Transport as a Probe of MutY/DNA Interaction

Boon, Elizabeth M. and Pope, Mary Ann and Williams, Scott D. and David, Sheila S. and Barton, Jacqueline K. (2002) DNA-Mediated Charge Transport as a Probe of MutY/DNA Interaction. Biochemistry, 41 (26). pp. 8464-8470. ISSN 0006-2960. doi:10.1021/bi012068c.

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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.

Item Type:Article
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URLURL TypeDescription
Boon, Elizabeth M.0000-0003-1891-839X
David, Sheila S.0000-0001-5873-7935
Barton, Jacqueline K.0000-0001-9883-1600
Additional Information:© 2002 American Chemical Society. Received November 19, 2001; Revised Manuscript Received March 13, 2002. Support from the National Institutes of Health (GM49216) to J.K.B. and (CA67985) to S.S.D. is gratefully acknowledged. We thank the members of the David Laboratory that purified the wild type MutY used in these experiments.
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Issue or Number:26
Record Number:CaltechAUTHORS:20160210-160315353
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Official Citation:DNA-Mediated Charge Transport as a Probe of MutY/DNA Interaction Elizabeth M. Boon, Mary Ann Pope, Scott D. Williams, Sheila S. David, and Jacqueline K. Barton Biochemistry 2002 41 (26), 8464-8470 DOI: 10.1021/bi012068c
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:64395
Deposited By: George Porter
Deposited On:19 Feb 2016 17:56
Last Modified:10 Nov 2021 23:30

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