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Effects of a hairpin polyamide on DNA melting: comparison with distamycin and Hoechst 33258

James, PL; Le Strat, L; Ellervik, Ulf LU ; Bratwall, C; Norden, B; Brown, T and Fox, KR (2004) In Biophysical Chemistry 111(3). p.205-212
Abstract
We have used DNase I footprinting and fluorescence melting studies to study the interaction of the hairpin polyamide Im-Py-Py-Py-(R)(H2N)gamma-Im`-PY-PY-PY-beta-Dp with its preferred binding sites (5'-WGWWCW; W=A or T) and other sequences. DNase I footprinting confirmed that the ligand binds to the sequence AGAACA at nanomolar concentrations and that changing the terminal A to G causes a dramatic decrease in affinity, while there was no. interaction with the reverse sequence WCWWGW Fluorescence melting studies with I I mer duplexes showed that the polyamide had very different effects on the forward (TGWWCT) and reverse (TCTAGT) sequences. At low concentrations, the polyamide produced biphasic melting curves with TGATCT, TGTACT and TGAACT,... (More)
We have used DNase I footprinting and fluorescence melting studies to study the interaction of the hairpin polyamide Im-Py-Py-Py-(R)(H2N)gamma-Im`-PY-PY-PY-beta-Dp with its preferred binding sites (5'-WGWWCW; W=A or T) and other sequences. DNase I footprinting confirmed that the ligand binds to the sequence AGAACA at nanomolar concentrations and that changing the terminal A to G causes a dramatic decrease in affinity, while there was no. interaction with the reverse sequence WCWWGW Fluorescence melting studies with I I mer duplexes showed that the polyamide had very different effects on the forward (TGWWCT) and reverse (TCTAGT) sequences. At low concentrations, the polyamide produced biphasic melting curves with TGATCT, TGTACT and TGAACT, suggesting a strong interaction. In contrast, the melting profiles with TCTAGT were always monophasic and showed much smaller concentration dependent changes in T-m. The polyamide also showed weak binding to the sequence TGATCT when one of the central AT pairs was replaced with an AC mismatch. These melting profiles were compared with those produced by the AT-selective minor groove binding agents distamycin and Hoechst 33258 at the same site's and at similar sequences containing A(5) and (AT)(3), which are expected to bind distamycin in the 1:1 and 2:1 modes, respectively. These ligands produced simple monophasic melting curves in which the T-m steadily increased as the ligand concentration was raised. (C) 2004 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Polyamide, Distamycin, Hoechst 33258, Molecular beacons, DNA melting
in
Biophysical Chemistry
volume
111
issue
3
pages
205 - 212
publisher
Elsevier
external identifiers
  • wos:000224852200003
  • pmid:15501563
  • scopus:6344221521
ISSN
1873-4200
DOI
10.1016/j.bpc.2004.06.001
language
English
LU publication?
yes
id
f3a8e924-2a51-42ee-b723-866565a961e1 (old id 141145)
date added to LUP
2007-07-10 11:24:24
date last changed
2017-01-01 07:19:02
@article{f3a8e924-2a51-42ee-b723-866565a961e1,
  abstract     = {We have used DNase I footprinting and fluorescence melting studies to study the interaction of the hairpin polyamide Im-Py-Py-Py-(R)(H2N)gamma-Im`-PY-PY-PY-beta-Dp with its preferred binding sites (5'-WGWWCW; W=A or T) and other sequences. DNase I footprinting confirmed that the ligand binds to the sequence AGAACA at nanomolar concentrations and that changing the terminal A to G causes a dramatic decrease in affinity, while there was no. interaction with the reverse sequence WCWWGW Fluorescence melting studies with I I mer duplexes showed that the polyamide had very different effects on the forward (TGWWCT) and reverse (TCTAGT) sequences. At low concentrations, the polyamide produced biphasic melting curves with TGATCT, TGTACT and TGAACT, suggesting a strong interaction. In contrast, the melting profiles with TCTAGT were always monophasic and showed much smaller concentration dependent changes in T-m. The polyamide also showed weak binding to the sequence TGATCT when one of the central AT pairs was replaced with an AC mismatch. These melting profiles were compared with those produced by the AT-selective minor groove binding agents distamycin and Hoechst 33258 at the same site's and at similar sequences containing A(5) and (AT)(3), which are expected to bind distamycin in the 1:1 and 2:1 modes, respectively. These ligands produced simple monophasic melting curves in which the T-m steadily increased as the ligand concentration was raised. (C) 2004 Elsevier B.V. All rights reserved.},
  author       = {James, PL and Le Strat, L and Ellervik, Ulf and Bratwall, C and Norden, B and Brown, T and Fox, KR},
  issn         = {1873-4200},
  keyword      = {Polyamide,Distamycin,Hoechst 33258,Molecular beacons,DNA melting},
  language     = {eng},
  number       = {3},
  pages        = {205--212},
  publisher    = {Elsevier},
  series       = {Biophysical Chemistry},
  title        = {Effects of a hairpin polyamide on DNA melting: comparison with distamycin and Hoechst 33258},
  url          = {http://dx.doi.org/10.1016/j.bpc.2004.06.001},
  volume       = {111},
  year         = {2004},
}