Visualising the dissociation of sequence selective ligands from individual binding sites on DNA
Visualising the dissociation of sequence selective ligands from individual binding sites on DNA
We have used a modification of the footprinting technique to measure the dissociation of mithramycin, echinomycin and nogalamycin from their binding sites in a natural DNA fragment. Complexes with radiolabelled DNA were dissociated by addition of unlabelled DIVA. Samples were removed at various times and subjected to DNase I digestion, and the rate of dissociation from each site was estimated from the time-dependent disappearance of the footprints. For echinomycin the slowest rate of dissociation is from ACGT, while the slowest site for mithramycin contains four contiguous guanines. The dissociation of nogalamycin is extremely slow, even from its weaker sites; the slowest rate was from ACGTA, which took longer than 4 h, even at 37°C.
Dissociation, Echinomycin, Mithramycin, Nogalamycin
118-122
Fletcher, Michael C.
81b0f822-354c-4e8a-a3da-e0ead19c1e90
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
12 February 1996
Fletcher, Michael C.
81b0f822-354c-4e8a-a3da-e0ead19c1e90
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
Fletcher, Michael C. and Fox, Keith R.
(1996)
Visualising the dissociation of sequence selective ligands from individual binding sites on DNA.
FEBS Letters, 380 (1-2), .
(doi:10.1016/0014-5793(96)00039-7).
Abstract
We have used a modification of the footprinting technique to measure the dissociation of mithramycin, echinomycin and nogalamycin from their binding sites in a natural DNA fragment. Complexes with radiolabelled DNA were dissociated by addition of unlabelled DIVA. Samples were removed at various times and subjected to DNase I digestion, and the rate of dissociation from each site was estimated from the time-dependent disappearance of the footprints. For echinomycin the slowest rate of dissociation is from ACGT, while the slowest site for mithramycin contains four contiguous guanines. The dissociation of nogalamycin is extremely slow, even from its weaker sites; the slowest rate was from ACGTA, which took longer than 4 h, even at 37°C.
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Published date: 12 February 1996
Keywords:
Dissociation, Echinomycin, Mithramycin, Nogalamycin
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Local EPrints ID: 475872
URI: http://eprints.soton.ac.uk/id/eprint/475872
ISSN: 0014-5793
PURE UUID: 0b5bf9be-2208-403d-84e3-57aa3fe34119
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Date deposited: 29 Mar 2023 16:47
Last modified: 17 Mar 2024 02:34
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Author:
Michael C. Fletcher
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