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  Protein-free parallel triple-stranded DNA complex formation

Shchyolkina, A. K., Timofeev, E. N., Lysov, Y. P., Florentiev, V. L., Jovin, T. M., & Arndt-Jovin, D. J. (2001). Protein-free parallel triple-stranded DNA complex formation. Nucleic Acids Research, 29, 986-995.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-F715-B Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-E6AD-3
Genre: Journal Article

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Shchyolkina, A. K., Author
Timofeev, E. N., Author
Lysov, Y. P., Author
Florentiev, V. L., Author
Jovin, T. M.1, Author              
Arndt-Jovin, D. J.1, Author              
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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 Abstract: A 14 nt DNA sequence 5'-AGAATGTGGCAAAG-3' from the zinc finger repeat of the human KRAB zinc finger protein gene ZNF91 bearing the intercalator 2-methoxy,6-chloro,9-amino acridine (Acr) attached to the sugar-phosphate backbone in various positions has been shown to form a specific triple helix (triplex) with a 16 bp hairpin (intramolecular) or a two-stranded (intermolecular) duplex having the identical sequence in:the same (parallel) orientation, Intra-molecular targets with the identical sequence in the antiparallel orientation and a non-specific target sequence were tested as controls, Apparent binding constants for formation of the tripler were determined by quantitating electrophoretic band shifts, Binding of the single-stranded oligonucleotide probe sequence to the target-led to an increase in the fluorescence anisotropy of acridine, The parallel orientation of the two Identical sequence segments was confirmed by measurement of fluorescence resonance energy transfer between the acridine on the 5'-end of the probe strand as donor and BODIPY-Texas Red on the 3'-amino group of either strand of the target duplex as acceptor, There was full protection from OsO4-bipyridine modification of thymines in the probe strand of the tripler, in accordance with the presumed tripler formation,which excluded displacement of the homologous duplex strand by the probe-intercalator conjugate, The implications of these results for the existence of protein-independent parallel triplexes are discussed.

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Language(s): eng - English
 Dates: 2005-08-052001
 Publication Status: Published in print
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 Identifiers: eDoc: 236892
Other: 34687
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Title: Nucleic Acids Research
Source Genre: Journal
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Pages: - Volume / Issue: 29 Sequence Number: - Start / End Page: 986 - 995 Identifier: -