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Binding of Hoechst with nucleic acids using fluorescence spectroscopy

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DOI: 10.4236/jbpc.2011.24052    5,251 Downloads   9,883 Views   Citations
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ABSTRACT

It has been shown that polarity of environment around Hoechst 33342 is almost unchanged while sorption of this fluorescent dye on a surface of the hairpin oligonucleotide HP1, t-RNA and DNA. At small concentrations, this dye, adsorbed on the surface of DNA, RNA or HP1, does not show any specificity to certain nucleotides. In the case of unwound sites of DNA or HP1, it can bind inside, but without the intercalation stacking with nucleotides. The energy transfer from nucleotide chromophores to Hoechst is absent due to their remoteness and also “bad” (non-stacking) orientation. The mutual fluorescence quenching of Hoechst by actinomycin D (AMD) and, vice versa, of 7-amino-actinomycin D (7AAMD) by Hoechst in DNA and HP1 is observed. It is due to dynamic deactivation and mutual replacing in binding sites.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Vekshin, N. (2011) Binding of Hoechst with nucleic acids using fluorescence spectroscopy. Journal of Biophysical Chemistry, 2, 443-447. doi: 10.4236/jbpc.2011.24052.

References

[1] Chen, F.M., Sha, F. and Chin, K.H. (2004) The nature of actinomycin D binding to d(AACCAXYG) sequence mo- tif. Nucleic Acids Research, 32, 271-277. doi:10.1093/nar/gkh178
[2] Vekshin, N., Savintsev, I., Kovalev, A., Yelemessov, R. and Wadkins, R. (2001) Solvatochromism of the excita- tion and emission spectra of 7-amino-actinomycin D: Im- plications for drug recognition of DNA secondary struc- tures. The Journal of Physical Chemistry: B, 105, 8461- 8467. doi:10.1021/jp011168p
[3] Vekshin, N.L. and Kovalev, A.E. (2006) Prompt non- stacking binding of actinomycin D to hairpin oligonu- cleotide HP1 and slow redistribution from HP1 to DNA. The Journal of Biochemistry, 140, 185-191. doi:10.1093/jb/mvj150
[4] Bitehtina, M.A. and Vekshin, N.L. (2008) 7-amino-ac- tinomycin as a fluorescent probe to un-twisting and de- naturation of DNA. Bioorganicheskaya Kchimiya, 34, 781-785.
[5] Vekshin, N.L. (2011) Biophysics of DNA-antibiotic com- plexes. Nova Press Inc, New York.
[6] Van Zant, G. and Fry, C.G. (1983) Hoechst 33342 of mouse bone marrow: Effects on colony-forming cells. Cytometry, 4, 40-46. doi:10.1002/cyto.990040106
[7] Sriram, M., Van der Marel, G.A., Roelen, H.L., Van Boom, J.H. and Wang, A.H. (1992) Conformation of B-DNA containing O6-ethyl-G-C base pairs stabilized by minor groove binding drugs: Molecular structure of d(CGC- [e6G]AATTCGCG complexed with Hoechst 33258 or Hoechst 33342. The EMBO Journal, 11, 225-232.
[8] Murata, S., Herman, P. and Lakowicz, J.R. (2001) Tex- ture analysis of fluorescence lifetime images of AT- and GC-rich regions in nuclei. Journal of Histochemistry and Cytochemistry, 49, 1443-1452. doi:10.1177/002215540104901112
[9] Adhikary, A., Buschmann, V., Muller, Ch. and Sauer, M. (2003) Ensemble and single-molecule fluorescence spec- troscopic study of the binding modes of the bis-benzi- midazole derivative Hoechst 33258 with DNA. Nucleic Acids Research, 31, 2178-2186. doi:10.1093/nar/gkg308
[10] Saito, M., Kobayashi, M., Iwabuchini, S.I., Motita, Y., Takamura, Y. and Tamiya, E. (2004) DNA condensation monitoring after interaction with Hoechst 33258 by ato- mic force microscopy and fluorescence spectroscopy. The Journal of Biochemistry, 36, 813-823. doi:10.1093/jb/mvh191
[11] Zimmer, C. and Wahnert, U. (1986) Nonintercalating DNA-binding ligands: Specificity of the interaction and their use as tools in biophysical, biochemical and biological investigations of the genetic material. Progress in Biophysics and Molecular Biology, 47, 31-112. doi:10.1016/0079-6107(86)90005-2
[12] Vekshin, N.L. (2009) Fluorescence spectroscopy of bio- macromolecules. Photon-vek, Pushchino.

  
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