RNA targeting by DNA binding drugs: structural, conformational and energetic aspects of the binding of quinacrine and DAPI to A-form and H(L)-form of poly(rC).poly(rG).
A key step in the rational design of new RNA binding small molecules necessitates a complete elucidation of the molecular aspects of the binding of existing molecules to RNA structures. This work focuses towards the understanding of the interaction of a DNA intercalator, quinacrine and a minor groove binder 4',6-diamidino-2-phenylindole (DAPI) with the right handed Watson-Crick base paired A-form and the left-handed Hoogsteen base paired H(L)-form of poly(rC).poly(rG) evaluated by multifaceted spectroscopic and viscometric techniques. The energetics of their interaction has also been elucidated by isothermal titration calorimetry. Results of this study converge to suggest that (i) quinacrine intercalates to both A-form and H(L)-form of poly(rC).poly(rG); (ii) DAPI shows both intercalative and groove-binding modes to the A-form of the RNA but binds by intercalative mode to the H(L)-form. Isothermal calorimetric patterns of quinacrine binding to both the forms of RNA and of DAPI binding to the H(L)-form are indicative of single binding while the binding of DAPI to the A-form reveals two kinds of binding. The binding of both the drugs to both conformations of RNA is exothermic; while the binding of quinacrine to both conformations and DAPI to the A-form (first site) is entropy driven, the binding of DAPI to the second site of A-form and H(L)-conformation is enthalpy driven. Temperature dependence of the binding enthalpy revealed that the RNA-ligand interaction reactions are accompanied by small heat capacity changes that are nonetheless significant. We conclude that the binding affinity characteristics and energetics of interaction of these DNA binding molecules to the RNA conformations are significantly different and may serve as data for the development of effective structure selective RNA-based antiviral drugs.
Sinha
Rangana
R
Biophysical Chemistry Laboratory, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Kolkata, India.
Hossain
Maidul
M
Kumar
Gopinatha Suresh
GS
D001665:Binding Sites; D002151:Calorimetry / Q000379:methods; D004247:DNA / Q000378:metabolism*; D007211:Indoles / Q000378:metabolism / Q000494:pharmacology*; D008968:Molecular Conformation; D011066:Poly C / Q000378:metabolism*; D011068:Poly G / Q000378:metabolism*; D011796:Quinacrine / Q000378:metabolism / Q000494:pharmacology*; D012313:RNA / Q000187:drug effects*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D007211:Indoles; D011068:Poly G; D011066:Poly C; C007293:DAPI; D012313:RNA; D004247:DNA; D011796:Quinacrine