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Zhou, HJ; Zhang, Y; Ou-Yang, ZC; Lindsay, SM; Feng, XZ; Balagurumoorthy, P; Harrington, RE; Harrington, RE , Arizona State Univ, Dept Microbiol, Tempe, AZ 85287 USA.
Conformation and rigidity of DNA microcircles containing waf1 response element for p53 regulatory protein
Source PublicationJOURNAL OF MOLECULAR BIOLOGY
KeywordBinding Domain Peptides Supercoiled Dna Gel-electrophoresis Torsional Rigidity Crystal-structure Core Domain Curved Dna Sequence Recognition Elasticity
AbstractThe tuner-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. (C) 2001 Academic Press.
2001
ISSN0022-2836
Volume306Issue:2Pages:227-238
Subject AreaPhysics
Indexed BySCI
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Cited Times:12[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.itp.ac.cn/handle/311006/12811
Collection理论物理所科研产出_SCI论文
Corresponding AuthorHarrington, RE , Arizona State Univ, Dept Microbiol, Tempe, AZ 85287 USA.
Recommended Citation
GB/T 7714
Zhou, HJ,Zhang, Y,Ou-Yang, ZC,et al. Conformation and rigidity of DNA microcircles containing waf1 response element for p53 regulatory protein[J]. JOURNAL OF MOLECULAR BIOLOGY,2001,306(2):227-238.
APA Zhou, HJ.,Zhang, Y.,Ou-Yang, ZC.,Lindsay, SM.,Feng, XZ.,...&Harrington, RE , Arizona State Univ, Dept Microbiol, Tempe, AZ 85287 USA..(2001).Conformation and rigidity of DNA microcircles containing waf1 response element for p53 regulatory protein.JOURNAL OF MOLECULAR BIOLOGY,306(2),227-238.
MLA Zhou, HJ,et al."Conformation and rigidity of DNA microcircles containing waf1 response element for p53 regulatory protein".JOURNAL OF MOLECULAR BIOLOGY 306.2(2001):227-238.
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