What makes thienoguanosine an outstanding fluorescent DNA probe?
Fiche publication
Date publication
septembre 2020
Journal
Journal of the American Chemical Society
Auteurs
Membres identifiés du Cancéropôle Est :
Pr MELY Yves, Dr RICHERT Ludovic, Pr DIDIER Pascal
Tous les auteurs :
Kuchlyan J, Martinez-Fernandez L, Mori M, Gavvala K, Ciaco S, Boudier C, Richert L, Didier P, Tor Y, Improta R, Mély Y
Lien Pubmed
Résumé
Thienoguanosine (thG) is an isomorphic guanosine (G) surrogate that almost perfectly mimics G in nucleic acids. To exploit its full potential and lay the foundation for future applications, twenty DNA duplexes, where the bases facing and neighboring thG were systematically varied, were thoroughly studied using fluorescence spectroscopy, molecular dynamics simulations and mixed quantum mechanical/molecular mechanics calculations, yielding a comprehensive understanding of its photophysics in DNA. In matched duplexes, thG's hypochromism was larger for flanking G/C residues but its fluorescence quantum yield (QY) and lifetime values were almost independent of the flanking bases. This was attributed to high duplex stability, which maintains a steady orientation and distance between nucleobases, so that a similar charge transfer (CT) mechanism governs the photophysics of thG independently of its flanking nucleobases. thG can therefore replace any G residue in matched duplexes, while always maintaining similar photophysical features. In contrast, the local destabilization induced by a mismatch or an abasic site restores a strong dependence of thG's QY and lifetime values on its environmental context, depending on the CT route efficiency and solvent exposure of thG. Due to this exquisite sensitivity, thG appears ideal for monitoring local structural changes and single nucleotide polymorphism. Moreover, thG's dominant fluorescence lifetime in DNA is unusually long (9-29 ns), facilitating its selective measurement in complex media using a lifetime-based or a time-gated detection scheme. Taken together, our data highlights thG as an outstanding emissive substitute for G with good QY, long fluorescence lifetimes and exquisite sensitivity to local structural changes.
Référence
J. Am. Chem. Soc.. 2020 Sep 11;: