Abstract:
:Fluorescence resonance energy transfer studies allow to determine global shape properties of nucleic acids and nucleoprotein complexes. In many DNA-protein complexes, the DNA is more or less bent and the degree of bending can be obtained by FRET. For example, the DNA in complex with the integration host factor (IHF) is kinked by approximately 160 degrees building a U-shaped structure. The two DNA helix ends come close to one another in space in a distance range easily measurable by FRET. The global DNA structure of this complex can be mimicked by introducing two regions with unpaired bases ('bulges') into the DNA each producing a sharp kink of approximately 80 degrees. These U-shaped DNA constructs were used to measure the electrostatic interaction of the two nearly parallel negatively charged DNA helix arms. The electrostatic repulsion between the helix arms, and as a consequence their distance, decreases with growing salt concentration of mono- or divalent cations. This experimental approach also allows the sensitive study of the local structure of DNA sequences positioned between the two bulges.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Lorenz M,Hillisch A,Diekmann Sdoi
10.1016/s1389-0352(01)00038-1subject
Has Abstractpub_date
2002-01-01 00:00:00pages
197-209issue
3eissn
0168-1656issn
1873-4863pii
S1389-0352(01)00038-1journal_volume
82pub_type
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