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Title: | Strand displacement of double-stranded DNA by triplex-forming antiparallel purine-hairpins |
Authors: | Coma, Sílvia; Eritja Casadellà, Ramón ![]() |
Keywords: | double stranded DNA DNA nucleic acid guanine purine pyrimidine derivative Base Sequence Electrophoretic Mobility Shift Assay Nucleic acid conformation Polymerase Chain Reaction |
Issue Date: | 2005 |
Publisher: | Mary Ann Liebert |
Citation: | Oligonucleotides |
Abstract: | We characterize the binding affinity and the thermodynamics of hybridization of triplex-forming antiparallel purine-hairpins composed of two antiparallel purine domains linked by a loop directed toward single-stranded and double-stranded DNA (ssDNA, dsDNA). Gel retardation assays and melting experiments reveal that a 13-mer purine-hairpin binds specifically and with a Kd of 8 × 10-8 M to polypyrimidine ssDNA to form a triple helical structure. Remarkably, we show that purine-hairpins also bind polypurine/polypyrimidine stretches included in a dsDNA of several hundred bp in length. Binding of purine-hairpins to dsDNA occurs by triplex formation with the polypyrimidine strand, causing displacement of the polypurine strand. Because triplex formation is restricted to polypurine/polypyrimidine stretches of dsDNA, we studied the triplex formation between purine-hairpins and polypyrimidine targets containing purine interruptions. We found that an 11-mer purine-hairpin with an adenine opposite to a guanine interruption in the polypyrimidine track binds to ssDNA and dsDNA, allowing expansion of the possible target sites and increase in the length of purine-hairpins. Thus, when using a 20-mer purine-hairpin targeting an interruption-containing polypyrimidine target, the binding affinity is increased compared to its 13-mer antiparallel purine-hairpin counterpart. Surprisingly, this increase is much more pronounced than that observed for a tail-clamp purine-hairpin extended up to 20 nt in the Watson-Crick domain only. Thus, triplex-forming antiparallel purine-hairpins can be a potentially useful strategy for both single-strand and double-strand nucleic acid recognition. |
Publisher version (URL): | DOI: 10.1089/oli.2005.15.269 |
URI: | http://hdl.handle.net/10261/124878 |
DOI: | http://dx.doi.org/10.1089/oli.2005.15.269 |
Appears in Collections: | (IQAC) Artículos |
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File | Description | Size | Format | |
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Oligonucleotides2005.pdf | Artículo principal | 1,81 MB | Adobe PDF | ![]() View/Open |
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