The Solution Conformation of a Trisdecanucleotide Containing the Consensus Binding Site of the dnaA Initiation Protein

Owen

2007

Eur Biophys J

A detailed analysis of the (13)C relaxation of (13)C-labelled spermine bound to duplex and quadruplex DNA is presented. T(1), T(2) and heteronuclear NOE data were collected at four (13)C frequencies (75.4, 125.7, 150.9 and 201.2 MHz). The data were analyzed in terms of a frequency-dependent order parameter, S (2)(omega), to estimate the generalized order parameter and the contributions to the relaxation from different motional frequencies in the picosecond-nanosecond timescale and from any exchange processes that may be occurring on the microsecond-millisecond timescale. The relaxation data was surprisingly similar for spermine bound to two different duplexes and a linear parallel quadruplex. Analysis of the relaxation data from these complexes confirmed the conclusions of previous studies that the dominant motion of spermine is independent of the macroscopic tumbling of the DNA and has an effective correlation time of approximately 50 ps. In contrast, spermine bound to a folded antiparallel quadruplex had faster relaxation rates, especially R (2). As with the other complexes, a fast internal motion of the order of 50 ps makes a substantial contribution to the relaxation. The generalized order parameter for spermine bound to duplex DNA and the linear quadruplex is small but is larger for spermine bound to the folded quadruplex. In the latter case, there is evidence for exchange between at least two populations of spermine occurring on the microsecond-millisecond timescale.

An investigation of the dynamics of spermine bound to duplex and quadruplex DNA by 13C NMR spectroscopy

Owen

2007

Eur Biophys J

A comparison of the association of spermine with duplex and quadruplex DNA by NMR

2003

FEBS Letters

The association of [1P P,1Q Q-13 C 2 ]spermine ([1P P,1Q Q-13 C 2 ]-N,NP P-bis(3-aminopropyl)-1,4-butanediamine) with duplex and quadruplex DNA has been studied by nuclear magnetic resonance spectroscopy. 1D NOESY experiments using two-way selective cross-polarization (ISI-SCP-NOESY) showed spermine intramolecular NOEs are either weakly positive or weakly negative when spermine is complexed to duplex B-DNA and linear four-stranded quadruplex DNA. In contrast, large negative intramolecular NOEs are observed when spermine is complexed to two distinct forms of folded quadruplex DNA suggesting greater immobilization of spermine on these folded DNA quadruplexes. No changes in the quadruplex stem structure are observed but there are minor changes to the loop structure of a two-stranded folded quadruplex on binding spermine. ß

Solution Structure of the Na⁺ form of the Dimeric Guanine Quadruplex [d(G₃T₄G3)]₂

European Journal of Biochemistry

Strahan

Owen

et al. 1995

The solution structure of the DNA quadruplex formed by the association of two strands of the DNA oligonucleotide, d(G3T4G3), in NaCl solution has been determined by 1H two‐dimensional NMR techniques, full relaxation matrix calculations and restrained molecular dynamics. The refined structure incorporates the sequences 5′‐GlsG2AG3AT4AT5AT6AT7AG8sG9AG10A‐3′ and 5′‐G11sG12AG13AT14AT15A. T16AT17AG18sG19sG20A‐3′ (where S and A denote syn and anti, respectively) in a three‐quartet, diagonal‐looped structure that we [Strahan, G. D., Shafer, R. H. & Keniry, M. A. (1994) Nucleic Acids Res. 22, 5447–5455] and others [Smith, F. W., Lau, F. W. & Feigon, J. (1994) Proc. Natl Acad. Sci. USA 91, 10546–10550] have described. The loop structure is compact and incorporates many of the features found in duplex hairpin loops including base stacking, intraloop hydrogen bonding and extensive van der Waals' interactions. The first and third loop thymines stack over the outermost G‐quartet and are also associated by hydrogen bonding. The second and the fourth loop thymines fold inwards in order to enhance van der Waals' interactions. The unexpected sequential syn‐syn deoxyguanosines in the quadruplex stem appear to be a direct consequence of the way DNA oligonucleotides fold and the subsequent search for the most stable loop structure. The implications of loop sequence and length on the structure of quadruplexes are discussed.