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Light‐activated cleavage of DNA by cobalt‐bleomycin

Light‐activated cleavage of DNA by cobalt‐bleomycin
Light‐activated cleavage of DNA by cobalt‐bleomycin

We have studied the light‐activated cleavage of DNA by cobalt‐bleomycin using a series of synthetic DNA fragments containing (AT)n and (GC)n. This cleavage reaction requires high concentrations of the antibiotic and appears to be a stoichiometric process rather than a catalytic process. We find that, in common with the iron‐complex, cobalt‐bleomycin can cleave at ApT steps within regions of alternating AT residues; ApT steps within other sequences including (AAT)n· (ATT)n are not good substrates for cobalt‐bleomycin cleavage. Some repetitive regions display an alternating pattern of cleavage products, revealing the preferred arrangement of ligand molecules along a saturated DNA lattice. A similar repetitive pattern is found for diethylpyrocarbonate modification and hydroxyl‐radical cleavage. Although cleavage of ApT and GpC proceeds at equivalent rates, the data suggest that bleomycin binds more tightly to the latter. Adenine residues on the 3′ side of both GpC‐cleavage and ApT‐cleavage sites are rendered more reactive to diethylpyrocarbonate, consistent with a ligand‐induced alteration in local DNA structure. The cobalt‐bleomycin‐binding site consists of not more than four base pairs, and may be as small as three base pairs.

0014-2956
173-181
NIGHTINGALE, Karl P.
a3a1d01a-fcb3-4877-9752-4b0f047b6779
FOX, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
NIGHTINGALE, Karl P.
a3a1d01a-fcb3-4877-9752-4b0f047b6779
FOX, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f

NIGHTINGALE, Karl P. and FOX, Keith R. (1994) Light‐activated cleavage of DNA by cobalt‐bleomycin. European Journal of Biochemistry, 220 (1), 173-181. (doi:10.1111/j.1432-1033.1994.tb18612.x).

Record type: Article

Abstract

We have studied the light‐activated cleavage of DNA by cobalt‐bleomycin using a series of synthetic DNA fragments containing (AT)n and (GC)n. This cleavage reaction requires high concentrations of the antibiotic and appears to be a stoichiometric process rather than a catalytic process. We find that, in common with the iron‐complex, cobalt‐bleomycin can cleave at ApT steps within regions of alternating AT residues; ApT steps within other sequences including (AAT)n· (ATT)n are not good substrates for cobalt‐bleomycin cleavage. Some repetitive regions display an alternating pattern of cleavage products, revealing the preferred arrangement of ligand molecules along a saturated DNA lattice. A similar repetitive pattern is found for diethylpyrocarbonate modification and hydroxyl‐radical cleavage. Although cleavage of ApT and GpC proceeds at equivalent rates, the data suggest that bleomycin binds more tightly to the latter. Adenine residues on the 3′ side of both GpC‐cleavage and ApT‐cleavage sites are rendered more reactive to diethylpyrocarbonate, consistent with a ligand‐induced alteration in local DNA structure. The cobalt‐bleomycin‐binding site consists of not more than four base pairs, and may be as small as three base pairs.

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Published date: February 1994

Identifiers

Local EPrints ID: 479229
URI: http://eprints.soton.ac.uk/id/eprint/479229
ISSN: 0014-2956
PURE UUID: 604df3da-7291-4b95-97b8-ffb47310737a
ORCID for Keith R. FOX: ORCID iD orcid.org/0000-0002-2925-7315

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Date deposited: 20 Jul 2023 16:46
Last modified: 17 Mar 2024 02:34

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Contributors

Author: Karl P. NIGHTINGALE
Author: Keith R. FOX ORCID iD

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