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题名: Conformation and rigidity of DNA microcircles containing waf1 response element for p53 regulatory protein
作者: Zhou, HJ ;  Zhang, Y ;  Ou-Yang, ZC ;  Lindsay, SM ;  Feng, XZ ;  Balagurumoorthy, P ;  Harrington, RE
刊名: JOURNAL OF MOLECULAR BIOLOGY
出版日期: 2001
卷号: 306, 期号:2, 页码:227-238
关键词: BINDING DOMAIN PEPTIDES ;  SUPERCOILED DNA ;  GEL-ELECTROPHORESIS ;  TORSIONAL RIGIDITY ;  CRYSTAL-STRUCTURE ;  CORE DOMAIN ;  CURVED DNA ;  SEQUENCE ;  RECOGNITION ;  ELASTICITY
学科分类: Physics
通讯作者: Harrington, RE , Arizona State Univ, Dept Microbiol, Tempe, AZ 85287 USA.
部门归属: Arizona State Univ, Dept Microbiol, Tempe, AZ 85287 USA; Chinese Acad Sci, Inst Theoret Phys, Beijing 100080, Peoples R China; Arizona State Univ, Dept Phys & Astron, Tempe, AZ 85287 USA
英文摘要: The 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.
收录类别: SCI
原文出处: 查看原文
WOS记录号: WOS:000167192400008
Citation statistics: 
内容类型: 期刊论文
URI标识: http://ir.itp.ac.cn/handle/311006/12811
Appears in Collections:理论物理所1978-2010年知识产出_期刊论文

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Recommended Citation:
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.
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