基于原子力显微镜研究p53蛋白与PBR322 DNA的相互作用

doi:10.3976/j.issn.1002-4026.2017.04.017

山东科学 ›› 2017, Vol. 30 ›› Issue (4): 106-111.doi: 10.3976/j.issn.1002-4026.2017.04.017

基于原子力显微镜研究p53蛋白与PBR322 DNA的相互作用

陈洋，王艳伟，杨光参*

温州大学物理与电子信息工程学院，浙江温州 325035

收稿日期:2017-03-22 出版日期:2017-08-20 发布日期:2017-08-20
通信作者: 杨光参。E-mail: yanggc@wzu.edu.cn E-mail:yanggc@wzu.edu.cn
作者简介:陈洋（1990—），男，硕士研究生，研究方向为生命现象中的凝聚态物理。E-mail: speedmatrix2006@126.com
基金资助:
国家自然科学基金(11274245, 10974146, 11304232)；温州大学创新基金（3162015033）

Study on interaction between p53 protein and PBR322 DNA based on atomic force microscopy

CHEN Yang, WANG Yan-wei, YANG Guang-can*

School of Physics and Electronic Information, Wenzhou University, Wenzhou, 325035, China

Received:2017-03-22 Online:2017-08-20 Published:2017-08-20

摘要/Abstract

摘要：

肿瘤（癌症）是由于细胞在复制过程中，DNA损伤不能修复导致细胞凋亡或者细胞无限增殖而形成的。DNA在细胞中转录和翻译都会涉及蛋白与DNA的结合，肿瘤抑制蛋白也是参与这一过程的关键蛋白之一。然而，众多研究发现，肿瘤抑制蛋白p53具有识别和修复损伤DNA的效果，对于细胞的凋亡、基因的保护和避免癌症发生有着重要的意义。有研究表明，金属镁离子和锌离子可以增强p53蛋白的结构稳定性和p53DNA的亲和力。因此，我们基于原子力显微镜（AFM），直观地呈现出p53蛋白与PBR322环状DNA相互作用的图像，同时发现p53蛋白可以使环状DNA自身形成聚集或者相交。但是，对于长度相当的5 000bp的线状DNA几乎没有这样的效果，而对于20 000bp DNA不会出现这样的现象。然而，在高浓度镁离子环境下，环状DNA会扭转成为麻花状，即形成超螺旋结构。这一现象，可为p53蛋白功能和作用机理研究提供指导，也为癌症治疗、癌症药物开发以及癌症检测方法提供启发。

关键词: 金属镁离子, PBR322环状DNA, DNA超螺旋, p53蛋白, 相互作用

Abstract:

Tumor (cancer) is caused by the apoptosis or proliferation of cells when DNA damage cannot be repaired in time in the process of replication. The processes of DNA transcription and expression are all involved in the interaction of protein and DNA in living cells. And, tumor suppressor protein is also one of the key proteins involved in this process. However, many studies have found that the tumor suppressor protein p53 can recognize and repair damagingDNA, which plays a significant role in cell apoptosis, gene protection and avoidance of cancer. Studies have shown that the structural stability of p53 protein and the affinity of p53DNA can be enhanced by metal ions, such as magnesium and zinc ions. Therefore, based on atomic force microscopy (AFM), the interaction image of p53 protein and PBR322 ring DNA is visually presented. And, it is found that p53 protein can promote a ring DNA aggregation or cross in itself. However, this phenomenon occurrence is rare for linear DNA with the length of 5 000bp, and it is not occurrence in 20 000bp DNA either. However, protein p53 can make ring DNA to be a twist shape in high magnesium concentration solution, that is, the supercoil shape is formed. This phenomenon may provide guidance for the study on the function and mechanism of protein p53, as well as provide insights into cancer therapy, cancer drug development and cancer detection methods.

Key words: PBR322 ring DNA, DNA supercoil, protein p53, interaction, metal magnesium ions

中图分类号:

陈洋，王艳伟，杨光参*. 基于原子力显微镜研究p53蛋白与PBR322 DNA的相互作用[J]. 山东科学, 2017, 30(4): 106-111.

CHEN Yang, WANG Yan-wei, YANG Guang-can* . Study on interaction between p53 protein and PBR322 DNA based on atomic force microscopy[J]. SHANDONG SCIENCE, 2017, 30(4): 106-111.

参考文献

［1］NUTTALL P, LEE K, CICCARELLA P, et al. Singlemolecule studies of unlabeled fulllength p53 protein binding to DNA［J］. Journal of Physical Chemistry B, 2016, 120(9)：21062114.
［2］ FUNK W D, PAK D T, KARAS R H, et al. A transcriptionally active DNAbinding site for human p53 protein complexes［J］. Molecular & Cellular Biology, 1992, 12(6): 28662871.
［3］ TOKINO T, THIAGALINGAM S, ELDEIRY W S, et al. p53 tagged sites from human genomic DNA［J］. Human Molecular Genetics, 1994, 3(9): 15371542.
［4］ JIAO Y, CHERNY D I, HEIM G, et al. Dynamic interactions of p53 with DNA in solution by timelapse atomic force microscopy［J］. Journal of Molecular Biology, 2001, 314(2): 233243.
［5］ VOGELSTEIN B, LANE D P, LEVINE A J. Surfing the TP53 network［J］. Nature, 2000, 408(6810): 307310.
［6］ BROWN C J, LAIN S, VERMA C S, et al. Awakening guardian angels: Drugging the p53 pathway［J］. Nature Reviews Cancer, 2009, 9(12): 86273.
［7］ LEVINE A J. p53, the cellular gatekeeper for growth and division［J］. Cell, 1997, 88(3): 323331.
［8］ HOLLSTEIN M, RICE K, GREENBLATT M S, et al. Database of p53 gene somatic mutations in human tumors and cell lines［J］. Nucleic Acids Research, 1994, 22(17): 35515.
［9］ HOLLSTEIN M, SHOMER B, GREENBLATT M, et al. Somatic point mutations in the p53 gene of human tumors and cell lines: updated compilation［J］. Nucleic Acids Research, 1996, 24(1): 1416.
［10］ NAGAICH A K, ZHURKIN V B, SAKAMOTO H, et al. Architectural accommodation in the complex of four p53 DNA binding domain peptides with the p21/waf1/cip1 DNA response element［J］. The Journal of biological chemistry, 1997, 272(23): 1483041.
［11］ WANG Y, REED M, WANG P, et al. p53 domains: identification and characterization of two autonomous DNAbinding regions［J］. Genes & Development, 1993, 7(12B): 2575.
［12］ WANG Y, SCHWEDES J F, PARKS D, et al. Interaction of p53 with its consensus DNAbinding site［J］. Molecular & Cellular Biology, 1995, 15(4): 21572165.
［13］ BALAGURUMOORTHY P, SAKAMOTO H, LEWIS M S, et al. Four p53 DNAbinding domain peptides bind natural p53response elements and bend the DNA［J］. Proceedings of the National Academy of Sciences, 1995, 92(19): 85915.
［14］ CHERNY D I, STRIKER G, SUBRAMANIAM V, et al. DNA bending due to specific p53 and p53 core domainDNA interactions visualized by electron microscopy［J］. Journal of Molecular Biology, 1999, 294(4): 10151026.
［15］ CAIN C, MILLER S, AHN J, et al. The N terminus of p53 regulates its dissociation from DNA［J］. Journal of Biological Chemistry, 2000, 275(51): 3994439953.
［16］ BARGONETTI J, MANFREDI J J, CHEN X, et al. A proteolytic fragment from the central region of p53 has marked sequencespecific DNAbinding activity when generated from wildtype but not from oncogenic mutant p53 protein［J］. Genes & Development, 1993, 7(12B): 2565.
［17］ BAKALKIN G, SELIVANOVA G, YAKOVLEVA T, et al. p53 binds singlestranded DNA ends through the Cterminal domain and internal DNA segments via the middle domain［J］. Nucleic Acids Research, 1995, 23(3): 362369.
［18］ RICHTER P H, EIGEN M. Diffusion controlled reaction rates in spheroidal geometry. Application to repressoroperator association and membrane bound enzymes［J］. Biophysical Chemistry, 1974, 2(2): 255263.
［19］ BERG O G, BLOMBERG C. Association kinetics with coupled diffusional flows. Special application to the lac repressoroperator system［J］. Biophysical Chemistry, 1976, 4(4): 367381.
［20］ BERG O G, BLOMBERG C. Association kinetics with coupled diffusion. An extension to coiledchain macromolecules applied to the lac repressoroperator system［J］. Biophysical Chemistry, 1977, 7(1): 33.
［21］ BERG O G, BLOMBERG C. Association kinetics with coupled diffusion : III. Ionicstrength dependence of the lac repressoroperator association［J］. Biophysical Chemistry, 1978, 8(4): 271280.
［22］ WINTER R B, VON HIPPEL P H. Diffusiondriven mechanisms of protein translocation on nucleic acids. 2. The Escherichia coli lac repressoroperator interaction: equilibrium measurements［J］. Biochemistry, 1981, 20(24): 69486960.
［23］张先恩. 生物结构自组装［J］. 科学通报, 2009, (18): 26822690.

［24］ OIJEN A M V. A singlemolecule characterization of P53 search on DNA［J］. Proceedings of the National Academy of Sciences of the United States of America, 2011, 108(2): 563568.
［25］ XUE Y, WANG S, FENG X. Influence of magnesium ion on the binding of p53 DNAbinding domain to DNAresponse elements［J］. Journal of Biochemistry, 2009, 146(1): 7785.
［26］ XUE Y, WANG S X. Effect of metal ion on the structural stability of tumour suppressor protein p53 DNAbinding domain［J］. Journal of Biochemistry, 2009, 146(2): 193200.
［27］ CASSINA V, SERUGGIA D, BERETTA G L, et al. Atomic force microscopy study of DNA conformation in the presence of drugs［J］. Biophysics of Structure & Mechanism, 2011, 40(1): 5968.
［28］ LIU Z, LI Z, ZHOU H, et al. Imaging DNA molecules on mica surface by atomic force microscopy in air and in liquid［J］. Microscopy Research & Technique, 2005, 66(4): 179185.
［29］ THUNDAT T, ALLISON D P, WARMACK R J. Stretched DNA structures observed with atomic force microscopy［J］. Nucleic Acids Research, 1994, 22(20): 42248.
［30］ TAFVIZI A, HUANG F, LEITH J S, et al. Tumor suppressor p53 slides on DNA with low friction and high stability［J］. Biophysical Journal, 2008, 95(1): L01L03.
［31］ LIU Y, GUTHOLD M, SNYDER M J, et al. AFM of selfassembled lambda DNAhistone networks［J］. Colloids & Surfaces B Biointerfaces, 2015, 134: 1725.

[1]	王娅妮, 王明锋. 原子系综在二次方相互作用演化下多体纠缠特性[J]. 山东科学, 2022, 35(2): 124-130.
[2]	师建茹,高耀,赵慧亮,田俊生,武嫣斐. 基于疏肝解郁功效的逍遥散寒苦肝和温辛肝性效关系研究[J]. 山东科学, 2022, 35(1): 37-44.

基于原子力显微镜研究p53蛋白与PBR322 DNA的相互作用

Study on interaction between p53 protein and PBR322 DNA based on atomic force microscopy

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

Metrics

本文评价

推荐阅读 0