Mg2+对Bloom综合症解旋酶与G4DNA结合的影响研究

华东师范大学学报(自然科学版) ›› 20120, Vol. 2012 ›› Issue (6): 81-88.doi:

Mg²⁺对Bloom综合症解旋酶与G4DNA结合的影响研究

骆衡^1-3，蔡明娟^1,2，陈祥^{1, 2}，丁玫^{1, 2}，李坤^1,2，许厚强^1,2

1.贵州大学动物科学学院高原山地动物遗传育种与繁殖省部共建教育部重点实验室，贵阳 550025； 2.贵州大学动物科学学院贵州省动物遗传育种与繁殖重点实验室，贵阳 550025；
3.贵州大学农学院贵州山地农业病虫害重点实验室，贵阳 550025

收稿日期:2011-09-01 修回日期:2011-12-01 出版日期:2012-11-25 发布日期:2013-01-11

Study on the effects of Mg²⁺ on the binding between Bloom syndrome helicase and G4DNA

LUO Heng ^1-3, CAI Ming-juan ^1,2, CHEN Xiang ^1,2, DING Mei ^1,2,LI Kun ^1,2, XU Hou-qiang ^1,2

1. Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, College of Animal Science, Guizhou University, Guiyang 550025, China; 2. Guizhou Key Laboratory of Animal Genetics, Breeding Reproduction, College of Animal Science, Guizhou University, Guiyang 550025, China; 3. The Provincial Key Laboratory for Agricultural Pest Management of Mountainous Region, Agricultural College, Guizhou University, Guiyang 550025, China

Received:2011-09-01 Revised:2011-12-01 Online:2012-11-25 Published:2013-01-11

摘要/Abstract

摘要： 利用荧光偏振技术检测了Mg²⁺对G4DNA、BLM-G4DNA复合物和BLM642-1290解旋酶与G4DNA结合的影响．结果表明，G4DNA荧光偏振值随着Mg²⁺浓度的增加而增加(〖WTBX〗P〖WTBZ〗<0.01)；BLM-G4DNA复合物的荧光偏振值随着Mg²⁺浓度的增加出现下降—升高—下降的变化趋势(〖WTBX〗P〖WTBZ〗<0.01)；G4DNA与BLM642-1290解旋酶结合的荧光偏振值随着Mg²⁺浓度的增加而逐渐下降(〖WTBX〗P〖WTBZ〗<0.01)；分析不同Mg²⁺浓度下两种分子结合的〖WTBX〗Kd〖WTBZ〗值，发现Mg²⁺浓度为3.0 mmol/L时，BLM642-1290解旋酶与G4DNA最容易结合，表明适量Mg²⁺浓度会促进BLM642-1290与G4DNA的结合，但会引起两种分子结合的形状、流动性和电荷等性质的改变．这些结果可为进一步研究BLM解旋酶对G4DNA的作用机理提供相关资料．

关键词: BLM解旋酶, G4DNA, Mg²⁺, 荧光偏振技术

Abstract: This paper studied the effects of Mg²⁺ on G4DNA, BLM-G4DNA complex, and the binding of BLM642-1290 helicase and G4DNA using fluorescence anisotropy technology.The results indicated that the fluorescence anisotropy of G4DNA increased with Mg²⁺concentration (〖WTBX〗P〖WTBZ〗<0.01); The anisotropy of BLM-G4DNA complex showed varied tendency of decrease-increase-decrease with Mg²⁺ concentration (〖WTBX〗P〖WTBZ〗<0.01)；The anisotropy of the binding between the helicase and G4DNA gradually declined following Mg²⁺ concentration (〖WTBX〗P〖WTBZ〗<0.01); The binding was most likely to occur between the helicase and G4DNA when Mg²⁺ concentrations was 3.0 mmol/L, which suggested that the suitable amount of Mg²⁺ could promote the binding of the helicase and G4DNA but that could lead to the change of the binding properties of two molecules including the shape, flow ability, and charge. These results provided useful data for studying the interaction mechanism between BLM helicase and G4DNA.

Key words: BLM helicase, G4DNA, Mg²⁺, fluorescence polarization technology

中图分类号:

R733.7

骆衡，蔡明娟，陈祥，丁玫，李坤，许厚强. Mg²⁺对Bloom综合症解旋酶与G4DNA结合的影响研究[J]. 华东师范大学学报(自然科学版), 20120, 2012(6): 81-88.

LUO Heng, CAI Ming-juan, CHEN Xiang, DING Mei,LI Kun, XU Hou-qiang. Study on the effects of Mg²⁺ on the binding between Bloom syndrome helicase and G4DNA[J]. Journal of East China Normal University(Natural Sc, 20120, 2012(6): 81-88.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: http://xblk.ecnu.edu.cn/CN/

http://xblk.ecnu.edu.cn/CN/Y20120/V2012/I6/81

参考文献

［1］ BACHRATI C Z, HICKSON I D. RecQ helicases: suppressors of tumorigenesis and premature aging［J］. Biochem J, 2003, 374(Pt 3): 577-606. 

［2］ MOUNIA A G. Bloom syndrome, genomic instability and cancer: the SOS-like hypothesis［J］. Cancer Letters, 2006, 236(1): 1-12. 

［3］ WONG I, MOORE K J M, BJOMSON K P, et al. ATPase Activity of 〖WTBX〗Escherichia coli〖WTBZ〗 Rep Helicase Is Dramatically Dependent on DNA Ligation and Protein Oligomeric States［J］. Biochemistry, 1996, 35(18): 5726-5734. 

［4］ PHAM X H, TUTEJIA N. Potent inhibition of DNA unwinding and ATPase activities of pea DNA helicase 45 by DNA-binding agents［J］. Biochem Biophys Res Comm, 2002, 294(2): 334-339.

［5］ LINDOR N M, FURUICHI Y, KITAO S, et al. Rothmund-Thomson syndrome due to RECQ4 helicase mutations: report and clinical and molecular comparisons with Bloom syndrome and Werner syndrome［J］. Am J Med Genet, 2000, 90(3): 223-228.

［6］ KITAO S, OHSUGI I, ICHIKAWA K, et al. Cloning of two new human helicase genes of the RecQ family: biological significance of multiple species in higher eukaryotes［J］. Genomics, 1998, 54(3): 443-452.

［7］ GERMAN J, ROE A M, LEPPERT M F, et al. Bloom syndrome: an analysis of consanguinous families assigns the locus mutated to chromosome band 15q26.1［J］. Proc Natl Acad Sci USA, 1994, 91(14): 6669-6673.

［8］ HICKSON I D. RecQ helicases: caretakers of the genome［J］. Nature Reviews Cancer, 2003(3): 169-178.

［9］ CHEOK C F, BACHRATI C Z, CHAN K L C, et al. Roles of the Bloom′s syndrome helicase in the maintenance of genome stability［J］. Biochemical Society Transactions, 2005, 33: 1456-1459. 

［10］ BACHRATI C Z, HICKSON I D. RecQ helicases: guardian angels of the DNA replication fork［J］. Chromosoma, 2008, 117(3): 219-233.

［11］ THOMPSON L H, SCHILD D. Recombinational DNA repair and human disease［J］. Mutat Res, 2002, 509(1-2): 49-78. 

［12］ XI X G. Helicases as antiviral and anticancer drug targets［J］. Current Medicinal Chemistry, 2007, 14(8): 503-517.

［13］ JANSCAK P, GARCIA P L, HAMBURGER F, et al. Characterization and mutational analysis of the RecQ core of the bloom syndrome protein［J］. J Mol Biol, 2005, 330: 29-42.

［14］殷菲, 刘建辉, 彭孝军. 以G-四链体为靶点的小分子端粒酶抑制剂研究进展［J］. 化学通报, 2004(4): 271-277.

［15］ YANG D Z, OKAMOTO K. Structural insights into G-quadruplexes: towards new anticancer drugs［J］. Future Med Chem, 2010, 2(4): 619-646.

［16］周江, 袁谷. 溶液pH 和阳离子对端粒G-四链体DNA 形成和结构的影响［J］. 化学通报, 2007, 65(16): 1728-1732.

［17］ DEMPSEY L A, SUN H, HANAKAHI L A, et al. G4 DNA binding by LR1 and its subunits, nucleolin and hnRNP D, a role for G-G pairing in immunoglobulin switch recombination［J］. The Journal of Biological Chemistry, 1999, 274(2): 1066-1071.

［18］胡晓文, 黄洪章. 端粒G-四联体—肿瘤治疗的新靶点［J］. 国外医学口腔医学分册, 2004, 31: 98-100.

［19］ HUPPERT J L, BALASUBRAMANIAN S. Prevalence of quadruplexes in the human genome［J］. Nucleic Acids Res, 2005, 33: 2908-2916.

［20］ HUPPERT J L, BALASUBRAMANIAN S. G-quadruplexes in promoters throughout the human genome［J］. Nucleic Acids Res, 2007, 35: 406-413.

［21］ SUN H, KAROW J K, HICKSON I D. The Bloom′s syndrome helicase unwinds G4 DNA［J］. The Journal of Biological Chemistry, 1998, 273, 42: 27587-27592.

［22］ GUO R B, RIGOLET P, ZARGARIAN L, et al. Structural and functional characterizations reveal the importance of a zinc binding domain in Bloom′s syndrome helicase［J］. Nucleic Acids Research, 2005, 33(10): 3109-3124.

［23］ CHEOK C F, BACHRATI C Z, CHAN K L, et al. Roles of the Bloom′s syndrome helicase in the maintenance of genome stability［J］.〖WTBX〗〖WTBZ〗Biochemical Society Transactions, 2005, 33(6): 1456-1459.

［24］ DOU S X, WANG P Y, XU H Q, et al. The DNA binding properties of the 〖WTBX〗Escherichia coli 〖WTBZ〗RecQ helicase［J］. J Biol Chem, 2004, 279(8): 6354-6363.

［25］ VERONIQUE L T, CATHERINE A R. Fluorescence anisotropy assays implicate protein-protein interactions in regulating trp repressor DNA binding［J］.Biochemistry, 1993, 32(30): 7753-7758.

［26］ LIU Y Y, WANG P Y, DOU S X, et al. Study the effects of divalent metallic ions on the combination of DNA and histones with fluorescence anisotropy assays［J］. Chinese Science Bulletin, 2007, 52(9): 1166-1171.

［27］ BJOMSON K P, AMARATUNGA M, MOORE K J, et al. Single-turnover kinetics of helicase-catalyzed DNA unwinding monitored continuously by fuorescence energy transfer［J］. Biochemistry, 1994, 33(47): 14306-14316.

［28］ RANEY K D, SOWERS L C, MILLAR D P, et al. A fuorescence-based assay for monitoring helicase activity［J］. Proc Natl Acad Sci USA, 1994, 91(14): 6644-6648.

［29］ DUGUID J, BLOOMFIELD V A, BENEVIDES J, et al. Raman spectroscopy of DNA-metal complexes. I. Interactions and conformational effects of the divalent cations: Mg, Ca, Sr, Ba, Mn, Co, Ni, Cu, Pd, and Cd［J］. Biophys J, 1993, 65: 1916-1928.

［30］ SUN H, JIANG Y J, YU Q S, et al. Molecular dynamics simulations on the role of structural Mg2+ ions in phosphoryl transfer catalyzed by GSK-3β［J］. Acta Phys-Chim Sin, 2011, 27(1): 207-212.

［31］骆衡, 陈祥, 段丽霞, 等. 大肠杆菌RecQ解旋酶的生物学活性分析［J］. 中国生物化学与分子生物学报, 2010, 26(12): 1143-1150.

［32］ CHEN X, LUO H, DUAN L X, et al. Effects of mercury on the structure and activity of BLM642-1290 helicase［J］. Biomedical and Environmental Sciences, 2011, 24(1): 47-55.

［33］ BERNSTEIN D A, ZITTEL M C, KECK J L. High-resolution structure of the E.coli RecQ helicase catalytic core［J］. EMBO J, 2003, 22(19): 4910-4921.

Mg²⁺对Bloom综合症解旋酶与G4DNA结合的影响研究

Study on the effects of Mg²⁺ on the binding between Bloom syndrome helicase and G4DNA

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

编辑推荐

Metrics

本文评价