Please use this identifier to cite or link to this item:
http://hdl.handle.net/2445/120404
Title: | Polypurine reverse-Hoogsteen (PPRH) oligonucleotides can form triplexes with their target sequences even under conditions where they fold into G-quadruplexes |
Author: | Solé Ferré, Anna Delagoutte, Emmanuelle Ciudad i Gómez, Carlos Julián Noé Mata, Verónica Alberti, Patrizia |
Keywords: | ADN Estructura molecular DNA Molecular structure |
Issue Date: | 9-Jan-2017 |
Publisher: | Nature Publishing Group |
Abstract: | Polypurine reverse-Hoogsteen (PPRH) oligonucleotides are non-modified DNA molecules composed of two mirror-symmetrical polypurine stretches linked by a five-thymidine loop. They can fold into reverse-Hoogsteen hairpins and bind to their polypyrimidine target sequence by Watson-Crick bonds forming a three-stranded structure. They have been successfully used to knockdown gene expression and to repair single-point mutations in cells. In this work, we provide an in vitro characterization (UV and fluorescence spectroscopy, gel electrophoresis and nuclease assays) of the structure and stability of two repair-PPRH oligonucleotides and of the complexes they form with their single-stranded targets. We show that one PPRH oligonucleotide forms a hairpin, while the other folds, in potassium, into a guanine-quadruplex (G4). However, the hairpin-prone oligonucleotide does not form a triplex with its single-stranded target, while the G4-prone oligonucleotide converts from a G4 into a reverse-Hoogsteen hairpin forming a triplex with its target sequence. Our work proves, in particular, that folding of a PPRH oligonucleotide into a G4 does not necessarily impair sequence-specific DNA recognition by triplex formation. It also illustrates an original example of DNA structural conversion of a G4 into a reverse-Hoogsteen hairpin driven by triplex formation; this kind of conversion might occur at particular loci of genomic DNA. |
Note: | Reproducció del document publicat a: https://doi.org/10.1038/srep39898 |
It is part of: | Scientific Reports, 2017, vol. 7 , num. 39898 |
URI: | http://hdl.handle.net/2445/120404 |
Related resource: | https://doi.org/10.1038/srep39898 |
ISSN: | 2045-2322 |
Appears in Collections: | Articles publicats en revistes (Bioquímica i Fisiologia) |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
670332.pdf | 1.38 MB | Adobe PDF | View/Open |
This item is licensed under a Creative Commons License