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DNA triple-helix formation on nucleosome core particles. Effect of length of the oligopurine tract

DNA triple-helix formation on nucleosome core particles. Effect of length of the oligopurine tract
DNA triple-helix formation on nucleosome core particles. Effect of length of the oligopurine tract

We have used DNase I footprinting to examine the formation of intermolecular triplexes on DNA fragments which have been complexed with nucleosome core particles. We have prepared five DNA fragments, based on the 160-bp tyrT sequence, which contain different length oligopurine tracts (up to 25 bp) at two different positions along the fragment, and have examined their availability for triple-helix formation after reconstituting onto nucleosome core particles. These results are compared with the formation of shorter triplexes in the same regions. In general we find that increasing the length of the complex does not facilitate nucleosomal triplex formation and that the most important factor affecting triplex formation is the position of the target site within the nucleosome-bound fragment. In some instances we find that longer oligonucleotides inhibit triplex formation. Although successful triplex formation was achieved on the longest nucleosome-bound oligopurine tracts, this was accompanied by changes in cleavage pattern that suggest oligonucleotide-induced changes in nucleosome structure.

Anti-gene, Footprinting, Nucleosomes, Sequence recognition, Triplex DNA
0014-2956
301-310
Brown, Philip M.
c910b8df-2849-4b26-8f69-3ca330836e9b
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
Brown, Philip M.
c910b8df-2849-4b26-8f69-3ca330836e9b
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f

Brown, Philip M. and Fox, Keith R. (1999) DNA triple-helix formation on nucleosome core particles. Effect of length of the oligopurine tract. European Journal of Biochemistry, 261 (1), 301-310. (doi:10.1046/j.1432-1327.1999.00279.x).

Record type: Article

Abstract

We have used DNase I footprinting to examine the formation of intermolecular triplexes on DNA fragments which have been complexed with nucleosome core particles. We have prepared five DNA fragments, based on the 160-bp tyrT sequence, which contain different length oligopurine tracts (up to 25 bp) at two different positions along the fragment, and have examined their availability for triple-helix formation after reconstituting onto nucleosome core particles. These results are compared with the formation of shorter triplexes in the same regions. In general we find that increasing the length of the complex does not facilitate nucleosomal triplex formation and that the most important factor affecting triplex formation is the position of the target site within the nucleosome-bound fragment. In some instances we find that longer oligonucleotides inhibit triplex formation. Although successful triplex formation was achieved on the longest nucleosome-bound oligopurine tracts, this was accompanied by changes in cleavage pattern that suggest oligonucleotide-induced changes in nucleosome structure.

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More information

Published date: 1 April 1999
Keywords: Anti-gene, Footprinting, Nucleosomes, Sequence recognition, Triplex DNA

Identifiers

Local EPrints ID: 476355
URI: http://eprints.soton.ac.uk/id/eprint/476355
ISSN: 0014-2956
PURE UUID: 09e5a51b-fe73-4df0-8f06-db3cbe974960
ORCID for Keith R. Fox: ORCID iD orcid.org/0000-0002-2925-7315

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Date deposited: 19 Apr 2023 16:46
Last modified: 17 Mar 2024 02:34

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Contributors

Author: Philip M. Brown
Author: Keith R. Fox ORCID iD

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