Title:
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Biophysical studies of the G-quadruplex motif
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The occurrence of multiple adjacent G-quadruplex-forming motifs in the
human genome, notably in the telomere, has promoted interest in how one
adjacently positioned quadruplex affects another. A range of oligonucleotides
were designed to feature one, two or three quadruplex-forming sequences
and their biophysical properties were compared. CD, DSC, thermal
denaturation experiments and supporting molecular simulations were the
main techniques used. Sequences were based on the human telomeric
sequence dAGGG(TTAGGG)3 and a parallel quadruplex-forming sequence
dTGGGTGGGTTTGGGTGGG.
Single and multiple telomeric quadruplexes formed similar topologies in
equivalent quantities. Decreased thermal stability was observed for tandem
quadruplexes. Divergence from a generally adequate two-state melting
process was attributed to interaction between quadruplex subunits and
domain-asymmetry. DSC analysis suggested that changes in heat capacity on
folding were non-trivial and that quadruplex stabilities based on Van't Hoff
enthalpy may be overestimated.
Melting behaviour showed that topology of adjacent quadruplexes affects
their interaction. Inter-quadruplex linker-length significantly affected
topology and could induce domain asymmetry with 5' and 3' quadruplexes
differing significantly in T m' Mixed telomeric and parallel quadruplex-forming
sequences suggested that 5' quadruplexes were significantly destabilised by a
3' partner. Steric hindrance was a possible explanation.
Quadruplex-binding ligands in the context of tandem quadruplexes were also
investigated. Acridine derivative, RHPS4, increased thermal stability and
induced topological changes in longer telomeric repeat sequences. A changed
pattern of thermal stabilities hinted at an indirect effect on mutual
quadruplex interactions via topological selection.
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