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. 1999 Aug 1;27(15):3018–3028. doi: 10.1093/nar/27.15.3018

The effect of sodium, potassium and ammonium ions on the conformation of the dimeric quadruplex formed by the Oxytricha nova telomere repeat oligonucleotide d(G(4)T(4)G(4)).

P Schultze 1, N V Hud 1, F W Smith 1, J Feigon 1
PMCID: PMC148525  PMID: 10454595

Abstract

The DNA sequence d(G(4)T(4)G(4)) [Oxy-1.5] consists of 1.5 units of the repeat in telomeres of Oxytricha nova and has been shown by NMR and X-ray crystallographic analysis to form a dimeric quadruplex structure with four guanine-quartets. However, the structure reported in the X-ray study has a fundamentally different conformation and folding topology compared to the solution structure. In order to elucidate the possible role of different counterions in this discrepancy and to investigate the conformational effects and dynamics of ion binding to G-quadruplex DNA, we compare results from further experiments using a variety of counterions, namely K(+), Na(+)and NH(4)(+). A detailed structure determination of Oxy-1.5 in solution in the presence of K(+)shows the same folding topology as previously reported with the same molecule in the presence of Na(+). Both conformations are symmetric dimeric quadruplexes with T(4)loops which span the diagonal of the end quartets. The stack of quartets shows only small differences in the presence of K(+)versus Na(+)counterions, but the T(4)loops adopt notably distinguishable conformations. Dynamic NMR analysis of the spectra of Oxy-1.5 in mixed Na(+)/K(+)solution reveals that there are at least three K(+)binding sites. Additional experiments in the presence of NH(4)(+)reveal the same topology and loop conformation as in the K(+)form and allow the direct localization of three central ions in the stack of quartets and further show that there are no specific NH(4)(+)binding sites in the T(4)loop. The location of bound NH(4)(+)with respect to the expected coordination sites for Na(+)binding provides a rationale for the difference observed for the structure of the T(4)loop in the Na(+)form, with respect to that observed for the K(+)and NH(4)(+)forms.

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Selected References

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  1. Blackburn E. H. Structure and function of telomeres. Nature. 1991 Apr 18;350(6319):569–573. doi: 10.1038/350569a0. [DOI] [PubMed] [Google Scholar]
  2. Blackburn E. H. Telomeres: structure and synthesis. J Biol Chem. 1990 Apr 15;265(11):5919–5921. [PubMed] [Google Scholar]
  3. Bouaziz S., Kettani A., Patel D. J. A K cation-induced conformational switch within a loop spanning segment of a DNA quadruplex containing G-G-G-C repeats. J Mol Biol. 1998 Sep 25;282(3):637–652. doi: 10.1006/jmbi.1998.2031. [DOI] [PubMed] [Google Scholar]
  4. Fang G., Cech T. R. Characterization of a G-quartet formation reaction promoted by the beta-subunit of the Oxytricha telomere-binding protein. Biochemistry. 1993 Nov 2;32(43):11646–11657. doi: 10.1021/bi00094a022. [DOI] [PubMed] [Google Scholar]
  5. Fang G., Cech T. R. The beta subunit of Oxytricha telomere-binding protein promotes G-quartet formation by telomeric DNA. Cell. 1993 Sep 10;74(5):875–885. doi: 10.1016/0092-8674(93)90467-5. [DOI] [PubMed] [Google Scholar]
  6. Fedoroff O. Y., Salazar M., Han H., Chemeris V. V., Kerwin S. M., Hurley L. H. NMR-Based model of a telomerase-inhibiting compound bound to G-quadruplex DNA. Biochemistry. 1998 Sep 8;37(36):12367–12374. doi: 10.1021/bi981330n. [DOI] [PubMed] [Google Scholar]
  7. Giraldo R., Rhodes D. The yeast telomere-binding protein RAP1 binds to and promotes the formation of DNA quadruplexes in telomeric DNA. EMBO J. 1994 May 15;13(10):2411–2420. doi: 10.1002/j.1460-2075.1994.tb06526.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hardin C. C., Henderson E., Watson T., Prosser J. K. Monovalent cation induced structural transitions in telomeric DNAs: G-DNA folding intermediates. Biochemistry. 1991 May 7;30(18):4460–4472. doi: 10.1021/bi00232a013. [DOI] [PubMed] [Google Scholar]
  9. Hud N. V., Schultze P., Sklenár V., Feigon J. Binding sites and dynamics of ammonium ions in a telomere repeat DNA quadruplex. J Mol Biol. 1999 Jan 8;285(1):233–243. doi: 10.1006/jmbi.1998.2327. [DOI] [PubMed] [Google Scholar]
  10. Hud N. V., Sklenár V., Feigon J. Localization of ammonium ions in the minor groove of DNA duplexes in solution and the origin of DNA A-tract bending. J Mol Biol. 1999 Feb 26;286(3):651–660. doi: 10.1006/jmbi.1998.2513. [DOI] [PubMed] [Google Scholar]
  11. Hud N. V., Smith F. W., Anet F. A., Feigon J. The selectivity for K+ versus Na+ in DNA quadruplexes is dominated by relative free energies of hydration: a thermodynamic analysis by 1H NMR. Biochemistry. 1996 Dec 3;35(48):15383–15390. doi: 10.1021/bi9620565. [DOI] [PubMed] [Google Scholar]
  12. Jing N., Hogan M. E. Structure-activity of tetrad-forming oligonucleotides as a potent anti-HIV therapeutic drug. J Biol Chem. 1998 Dec 25;273(52):34992–34999. doi: 10.1074/jbc.273.52.34992. [DOI] [PubMed] [Google Scholar]
  13. Kang C., Zhang X., Ratliff R., Moyzis R., Rich A. Crystal structure of four-stranded Oxytricha telomeric DNA. Nature. 1992 Mar 12;356(6365):126–131. doi: 10.1038/356126a0. [DOI] [PubMed] [Google Scholar]
  14. Keniry M. A., Strahan G. D., Owen E. A., Shafer R. H. Solution structure of the Na+ form of the dimeric guanine quadruplex [d(G3T4G3)]2. Eur J Biochem. 1995 Oct 15;233(2):631–643. doi: 10.1111/j.1432-1033.1995.631_2.x. [DOI] [PubMed] [Google Scholar]
  15. Laughlan G., Murchie A. I., Norman D. G., Moore M. H., Moody P. C., Lilley D. M., Luisi B. The high-resolution crystal structure of a parallel-stranded guanine tetraplex. Science. 1994 Jul 22;265(5171):520–524. doi: 10.1126/science.8036494. [DOI] [PubMed] [Google Scholar]
  16. Macaya R. F., Schultze P., Smith F. W., Roe J. A., Feigon J. Thrombin-binding DNA aptamer forms a unimolecular quadruplex structure in solution. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3745–3749. doi: 10.1073/pnas.90.8.3745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mazumder A., Neamati N., Ojwang J. O., Sunder S., Rando R. F., Pommier Y. Inhibition of the human immunodeficiency virus type 1 integrase by guanosine quartet structures. Biochemistry. 1996 Oct 29;35(43):13762–13771. doi: 10.1021/bi960541u. [DOI] [PubMed] [Google Scholar]
  18. Miura T., Benevides J. M., Thomas G. J., Jr A phase diagram for sodium and potassium ion control of polymorphism in telomeric DNA. J Mol Biol. 1995 Apr 28;248(2):233–238. doi: 10.1016/s0022-2836(95)80046-8. [DOI] [PubMed] [Google Scholar]
  19. Phillips K., Dauter Z., Murchie A. I., Lilley D. M., Luisi B. The crystal structure of a parallel-stranded guanine tetraplex at 0.95 A resolution. J Mol Biol. 1997 Oct 17;273(1):171–182. doi: 10.1006/jmbi.1997.1292. [DOI] [PubMed] [Google Scholar]
  20. Piotto M., Saudek V., Sklenár V. Gradient-tailored excitation for single-quantum NMR spectroscopy of aqueous solutions. J Biomol NMR. 1992 Nov;2(6):661–665. doi: 10.1007/BF02192855. [DOI] [PubMed] [Google Scholar]
  21. Schultze P., Feigon J. Chirality errors in nucleic acid structures. Nature. 1997 Jun 12;387(6634):668–668. doi: 10.1038/42632. [DOI] [PubMed] [Google Scholar]
  22. Schultze P., Smith F. W., Feigon J. Refined solution structure of the dimeric quadruplex formed from the Oxytricha telomeric oligonucleotide d(GGGGTTTTGGGG). Structure. 1994 Mar 15;2(3):221–233. doi: 10.1016/s0969-2126(00)00023-x. [DOI] [PubMed] [Google Scholar]
  23. Smith F. W., Feigon J. Quadruplex structure of Oxytricha telomeric DNA oligonucleotides. Nature. 1992 Mar 12;356(6365):164–168. doi: 10.1038/356164a0. [DOI] [PubMed] [Google Scholar]
  24. Smith F. W., Feigon J. Strand orientation in the DNA quadruplex formed from the Oxytricha telomere repeat oligonucleotide d(G4T4G4) in solution. Biochemistry. 1993 Aug 24;32(33):8682–8692. doi: 10.1021/bi00084a040. [DOI] [PubMed] [Google Scholar]
  25. Smith F. W., Lau F. W., Feigon J. d(G3T4G3) forms an asymmetric diagonally looped dimeric quadruplex with guanosine 5'-syn-syn-anti and 5'-syn-anti-anti N-glycosidic conformations. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10546–10550. doi: 10.1073/pnas.91.22.10546. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Strahan G. D., Keniry M. A., Shafer R. H. NMR structure refinement and dynamics of the K+-[d(G3T4G3)]2 quadruplex via particle mesh Ewald molecular dynamics simulations. Biophys J. 1998 Aug;75(2):968–981. doi: 10.1016/S0006-3495(98)77585-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sundquist W. I., Klug A. Telomeric DNA dimerizes by formation of guanine tetrads between hairpin loops. Nature. 1989 Dec 14;342(6251):825–829. doi: 10.1038/342825a0. [DOI] [PubMed] [Google Scholar]
  28. Wang K. Y., McCurdy S., Shea R. G., Swaminathan S., Bolton P. H. A DNA aptamer which binds to and inhibits thrombin exhibits a new structural motif for DNA. Biochemistry. 1993 Mar 2;32(8):1899–1904. doi: 10.1021/bi00059a003. [DOI] [PubMed] [Google Scholar]
  29. Williamson J. R. G-quartet structures in telomeric DNA. Annu Rev Biophys Biomol Struct. 1994;23:703–730. doi: 10.1146/annurev.bb.23.060194.003415. [DOI] [PubMed] [Google Scholar]
  30. Williamson J. R., Raghuraman M. K., Cech T. R. Monovalent cation-induced structure of telomeric DNA: the G-quartet model. Cell. 1989 Dec 1;59(5):871–880. doi: 10.1016/0092-8674(89)90610-7. [DOI] [PubMed] [Google Scholar]
  31. Wyatt J. R., Vickers T. A., Roberson J. L., Buckheit R. W., Jr, Klimkait T., DeBaets E., Davis P. W., Rayner B., Imbach J. L., Ecker D. J. Combinatorially selected guanosine-quartet structure is a potent inhibitor of human immunodeficiency virus envelope-mediated cell fusion. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1356–1360. doi: 10.1073/pnas.91.4.1356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zahler A. M., Williamson J. R., Cech T. R., Prescott D. M. Inhibition of telomerase by G-quartet DNA structures. Nature. 1991 Apr 25;350(6320):718–720. doi: 10.1038/350718a0. [DOI] [PubMed] [Google Scholar]
  33. Zakian V. A. Structure and function of telomeres. Annu Rev Genet. 1989;23:579–604. doi: 10.1146/annurev.ge.23.120189.003051. [DOI] [PubMed] [Google Scholar]

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