Interactions of nucleic acids with fluorescent dyes: spectral properties of condensed complexes.
Interaction of cations with nucleic acids (NA) often results in condensation of the product. The driving force of aromatic cation-induced condensation is the cooperative interaction between ligand and single-stranded (ss) NA. This type of reaction is highly specific with regard to the primary and secondary structure of NA, and results in destabilization of the latter. The spectral properties of fluorescent intercalating and non-intercalating ligands [acridine orange, pyronin Y(G), DAPI, Hoechst 33258, and Hoechst 33342]-NA complexes were studied in both the relaxed and condensed form. The changes in absorption, excitation, and fluorescence emission spectra and fluorescence yield that followed the condensation were examined. Although some of these effects can be explained by changes in solvation of the fluorophore and its interaction with NA bases and the solvent, the overall effect of condensation on spectral properties of the complex is unpredictable. In particular, no correlation was found between these effects and the ds DNA binding mode of these ligands. Nevertheless, the spectral data associated with polymer condensation can yield information about the composition and structure of NA and can explain some nonspecific interactions of these probes.
Laboratory of Experimental Cell Research, Sloan-Kettering Institute for Cancer Research, New York, New York 10021.
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D000165:Acridine Orange / Q000378:metabolism; D000818:Animals; D001562:Benzimidazoles / Q000378:metabolism; D006690:Bisbenzimidazole / Q000378:metabolism; D002417:Cattle; D002843:Chromatin / Q000032:analysis / Q000378:metabolism; D015342:DNA Probes; D005456:Fluorescent Dyes / Q000378:metabolism*; D007211:Indoles / Q000378:metabolism; D008163:Luminescent Measurements; D009696:Nucleic Acids / Q000378:metabolism*; D011754:Pyronine / Q000378:metabolism; D013053:Spectrophotometry; D014344:Trypanocidal Agents / Q000378:metabolism
D001562:Benzimidazoles; D002843:Chromatin; D015342:DNA Probes; D005456:Fluorescent Dyes; D007211:Indoles; D009696:Nucleic Acids; D014344:Trypanocidal Agents; C007293:DAPI; D000165:Acridine Orange; D006690:Bisbenzimidazole; C017807:bisbenzimide ethoxide trihydrochloride; D011754:Pyronine