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Title

Application of multivariate curve resolution to characterize conformational pH-transitions structures of i-motif DNA studied by time-resolved fluorescence spectroscopy

AuthorsBenabou, Sanae; Ruckebusch, Cyril; Sliwa, Michel; Aviñó, Anna ; Eritja Casadellà, Ramón ; Gargallo, Raimundo; de Juan, Anna
Issue Date11-Jun-2019
AbstractRecently, the presence of i-motif structures at C-rich sequences in human cells and their regulatory functions has been demonstrated. Despite numerous steady-state studies on i-motif at neutral and slightly acidic pH, the investigation of the detailed conformation of this biological structure using fluorescence spectroscopy at femtosecond time scale and the related decay curves and lifetimes has been scarcely reported. In this work, the fluorescence lifetime of labelled molecular beacon i- motif-forming DNA sequences at different pH values is studied. The influence of the nature of bases at the lateral loops, as well as the presence of a Watson-Crick-stabilized hairpin are studied by means of time-correlated single-photon counting (TCSPC) technique, which allows characterizing the existence of several conformers for which the fluorophore has lifetimes ranging from picosecond to nanosecond. The information on the existence of different i-motif structures at different pH values has been obtained by the combination of classical global decay fitting of fluorescence traces, which provides lifetimes associated with the events defined by the decay of each sequence, and multivariate analysis, such as Principal Component Analysis (PCA) or Multivariate Curve Resolution based on Alternating Least Squares (MCR-ALS). Multivariate analysis, which is seldom used for this kind of data, was crucial to explore similarities and differences of behaviour among the different DNA sequences and to model the presence and identity of the conformations involved in the pH range of interest.
URIhttp://hdl.handle.net/10261/203557
Appears in Collections:(IQAC) Comunicaciones congresos
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