Study on the effects of Mg<sup>2+</sup> on the binding between Bloom syndrome helicase and G4DNA

LUO Heng; CAI Ming-juan; CHEN Xiang; DING Mei; LI Kun; XU Hou-qiang; 

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Article Navigation > Journal of East China Normal University (Natural Sciences) > 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 Sciences), 2012, (6): 81-88.

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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 Sciences), 2012, (6): 81-88.

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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 Date: 2011-09-01
Rev Recd Date: 2011-12-01
Publish Date: 2012-11-25

Abstract

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.
- BLM helicase,
- G4DNA,
- Mg2+,
- fluorescence polarization technology

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References(1)

References

[1]

［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.

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