Publications (2)4 Total impact
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Article: Conformation and rigidity of DNA microcircles containing waf1 response element for p53 regulatory protein.
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ABSTRACT: The tumor-suppressor activity of p53 is closely related to its DNA-binding properties. It binds a number of DNA response-elements and it is likely that these share a common structural feature. Here, we present a new, general method to determine the absolute twist of flexible DNA promoter sequences based on direct imaging of the topology of microcircles containing the sequences. We have used magnetically driven dynamic force microscopy ("MacMode" AFM) to observe, in solution, the conformation of 168 base-pair DNA microcircles, each containing four equally spaced copies of the waf1/cip1/p21 p53 response-element. Analysis of the images showed that the microcircles are markedly puckered with a small excess of negatively writhed molecules. The average measured values of writhe are 0.109+/-0.013 (for 60 positively writhed molecules) and -0.098+/-0.011 (for 65 negatively writhed molecules). These values lead directly to a difference in linking number for the positively and negatively writhed molecules prior to ligation, from which we derive a twist mismatch of 178 degrees (overtwist). This is 44.5 degrees for each 42-mer precursor containing a single waf1/cip1/p21 p53 response-element, in good agreement with the range of values deduced by indirect biochemical techniques. The two values of writhe may also be used to determine the ratio of the bending (B) to twisting (C) rigidity, yielding B/C=0.23. This is about one-third of the value for long, random-sequence DNA, suggesting that the waf1/cip1/p21 p53 response-element is extremely flexible, a result that is also consistent with indirect biochemical experiments. These results support the idea, proposed by us earlier, that torsional stress may play a role in the regulation of p53 binding through modulation of twist at the binding site.Journal of Molecular Biology 03/2001; 306(2):227-38. · 4.00 Impact Factor -
Article: Elastic property of single double-stranded DNA molecules: theoretical study and comparison with experiments.
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ABSTRACT: This paper aims at a comprehensive understanding of the novel elastic property of double-stranded DNA (dsDNA) discovered very recently through single-molecule manipulation techniques. A general elastic model for double-stranded biopolymers is proposed, and a structural parameter called the folding angle straight phi is introduced to characterize their deformations. The mechanical property of long dsDNA molecules is then studied based on this model, where the base-stacking interactions between DNA adjacent nucleotide base pairs, the steric effects of base pairs, and the electrostatic interactions along DNA backbones are taken into account. Quantitative results are obtained by using a path integral method, and excellent agreement between theory and the observations reported by five major experimental groups are attained. The strong intensity of the base stacking interactions ensures the structural stability of DNA, while the short-ranged nature of such interactions makes externally stimulated large structural fluctuations possible. The entropic elasticity, highly extensibility, and supercoiling property of DNA are all closely related to this account. The present work also suggests the possibility that negative torque can induce structural transitions in highly extended DNA from the right-handed B form to left-handed configurations similar to the Z-form configuration. Some formulas concerned with the application of path integral methods to polymeric systems are listed in the Appendixes.Physical review. E, Statistical physics, plasmas, fluids, and related interdisciplinary topics 08/2000; 62(1 Pt B):1045-58.
