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Kinetic studies on the formation of DNA triplexes containing the nucleoside analogue 2'-O-(2-aminoethyl)-5-(3-amino-1-propynyl)uridine

Kinetic studies on the formation of DNA triplexes containing the nucleoside analogue 2'-O-(2-aminoethyl)-5-(3-amino-1-propynyl)uridine
Kinetic studies on the formation of DNA triplexes containing the nucleoside analogue 2'-O-(2-aminoethyl)-5-(3-amino-1-propynyl)uridine
We have examined the kinetics of triple helix formation of oligonucleotides that contain the nucleotide analogue 2'-O-(2-aminoethyl)-5-(3-amino-1-propynyl)uridine (bis-amino-U, BAU), which forms very stable base triplets with AT. Triplex stability is determined by both the number and location of the modifications. BAU-containing oligonucleotides generate triplexes with extremely slow kinetics, as evidenced by 14 °C hysteresis between annealing and melting profiles even when heated at a rate as slow as 0.2 °C min–1. The association kinetics were measured by analysis of the hysteresis profiles, temperature-jump relaxation and DNase I footprinting. We find that the slow kinetics are largely due to the decreased rate of dissociation; BAU modification has little effect on the association reaction. The sequence selectivity is also due to the slower dissociation of BAU from AT than other base pairs.
1477-0520
122-129
Rusling, David A.
d08f1f97-f8a9-4980-a025-ae41c23a938f
Broughton-Head, Victoria J.
d094aa77-c889-4747-84d2-55542805c67c
Tuck, Alex
38b69eb3-abc4-416c-9843-57dcb7f8d9c9
Khairallah, Hannah
8de8dfc6-a02b-42e7-8dca-36a9532cf992
Osborne, Sadie D.
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Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
Rusling, David A.
d08f1f97-f8a9-4980-a025-ae41c23a938f
Broughton-Head, Victoria J.
d094aa77-c889-4747-84d2-55542805c67c
Tuck, Alex
38b69eb3-abc4-416c-9843-57dcb7f8d9c9
Khairallah, Hannah
8de8dfc6-a02b-42e7-8dca-36a9532cf992
Osborne, Sadie D.
9777c83c-8998-4fd6-913e-04d3902ee9cd
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f

Rusling, David A., Broughton-Head, Victoria J., Tuck, Alex, Khairallah, Hannah, Osborne, Sadie D., Brown, Tom and Fox, Keith R. (2008) Kinetic studies on the formation of DNA triplexes containing the nucleoside analogue 2'-O-(2-aminoethyl)-5-(3-amino-1-propynyl)uridine. Organic & Biomolecular Chemistry, 6 (1), 122-129. (doi:10.1039/b713088k). (PMID:18075656)

Record type: Article

Abstract

We have examined the kinetics of triple helix formation of oligonucleotides that contain the nucleotide analogue 2'-O-(2-aminoethyl)-5-(3-amino-1-propynyl)uridine (bis-amino-U, BAU), which forms very stable base triplets with AT. Triplex stability is determined by both the number and location of the modifications. BAU-containing oligonucleotides generate triplexes with extremely slow kinetics, as evidenced by 14 °C hysteresis between annealing and melting profiles even when heated at a rate as slow as 0.2 °C min–1. The association kinetics were measured by analysis of the hysteresis profiles, temperature-jump relaxation and DNase I footprinting. We find that the slow kinetics are largely due to the decreased rate of dissociation; BAU modification has little effect on the association reaction. The sequence selectivity is also due to the slower dissociation of BAU from AT than other base pairs.

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e-pub ahead of print date: 1 November 2007
Published date: January 2008

Identifiers

Local EPrints ID: 142599
URI: http://eprints.soton.ac.uk/id/eprint/142599
ISSN: 1477-0520
PURE UUID: 473d6838-4925-43cf-88b1-8a0f73be69cb
ORCID for David A. Rusling: ORCID iD orcid.org/0000-0002-7442-686X
ORCID for Keith R. Fox: ORCID iD orcid.org/0000-0002-2925-7315

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Date deposited: 01 Apr 2010 16:02
Last modified: 14 Mar 2024 02:33

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Contributors

Author: David A. Rusling ORCID iD
Author: Victoria J. Broughton-Head
Author: Alex Tuck
Author: Hannah Khairallah
Author: Sadie D. Osborne
Author: Tom Brown
Author: Keith R. Fox ORCID iD

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