BLM解旋酶催化核心与G4DNA结合和解链特性

骆衡; 许厚强; 蔡明娟; 陈祥; 丁玫; 李坤; 孟惠惠

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The binding and unwinding properties of the Bloom helicase catalytic core to the G4DNA structure

首发时间：2012-03-22

LUO Heng ¹
LUO Heng, (1985-),male,Ph.D. Students,Molecular Biology of the Cell.
XU Houqiang ^{2
3}
XU Houqiang(1957),male,professor,Molecular Biology of the Cell
CAI Mingjuan ⁴ CHEN Xiang ⁴ DING Mei ⁴ LI Kun ⁴ MENG Huihui ⁴

1、Guizhou Key Laboratory of Animal Genetics and Breeding Reproduction,College of Animal Sciences,Guizhou University, GuiYang 550025
2、Provincial Key Laboratory for Agricultural Pest Management of Mountainous Region of Guizhou University
3、Key Laboratory of Ministry of Education for Green Pesticide and Agrobioengineering, Guizhou University
4、Guizhou Key Laboratory of Animal Genetics and Breeding Reproduction,College of Animal Sciences,Guizhou University

Abstract：G4DNA, which widely exists in the structure of the telomeres in normal cells, plays a pivotal part in the process of prolonging the telomere DNA by catalyzing the enzyme telomerase. Bloom (BLM) helicase, an important member of the RecQ DNA helicase family, plays an important role in DNA metabolism, including DNA replication, repair, transcription, and recombination. The unwinding of G4DNA requires DNA helicase participation, which is crucial for maintaining chromosomal stability. This study was conducted to determine the DNA-binding and unwinding properties of the BLM helicase catalytic core to G4DNA using fluorescence polarization and the electrophoretic mobility shift assay (EMSA). The results revealed that the BLM helicase catalytic core could bind and unwind G4DNA. The molecular affinity of G4DNA binding by the helicase was dependent on the single-stranded DNA (ssDNA) terminals in the G4DNA; the helicase binds to the G4DNA where one helicase monomer covers approximately 10 nucleotides at the 3' or 5' ssDNA tail. The unwinding of G4DNA was dependent on the presence of a 3' ssDNA tail and ATP; the G4DNA with only a 3' ssDNA tail was the most favorable substrate to be unwound by the BLM helicase catalytic core, and required 3' ssDNA tails of at least 10 nt in length for efficient unwinding. The blunt-ended G4DNA was loosely bound and partly unwound by the helicase catalytic core. The experimental results presented are beneficial to further the understanding the functionality of BLM helicase in cells.

keywords： Binding BLM helicase G4DNA unwinding fluorescence polarization

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BLM解旋酶催化核心与G4DNA结合和解链特性

骆衡 ¹
LUO Heng, (1985-),male,Ph.D. Students,Molecular Biology of the Cell.
许厚强 ²
XU Houqiang(1957),male,professor,Molecular Biology of the Cell
蔡明娟 ³ 陈祥 ⁴ 丁玫 ⁵ 李坤 ⁵ 孟惠惠 ⁵

1、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,贵州大学动物科学学院，贵阳 550025
2、贵州大学贵州省山地农业病虫害重点实验室
3、贵州大学教育部绿色农药与农业生物工程重点实验室
4、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,贵州大学动物科学学院
5、Guizhou Key Laboratory of Animal Genetics and Breeding Reproduction,College of Animal Sciences,Guizhou University

摘要：G4DNA 广泛存在于正常细胞的端粒DNA结构中，在通过端粒酶催化而延长端粒DNA的过程中起着重要作用。Bloom (BLM)解旋酶，是RecQ家族DNA解旋酶的重要成员，在DNA复制、修复、转录及重组等DNA代谢过程具有重要功能。G4DNA的解链需要DNA解旋酶的参与，这对于维持染色体的稳定性具有至关重要的作用。本研究利用荧光偏振技术和电泳迁移率实验(EMSA)研究了BLM解旋酶催化核心与G4DNA结合和解链的特性。结果表明：BLM解旋酶催化核心能够结合和解链G4DNA。BLM解旋酶催化核心和G4DNA结合的分子亲和力依赖于G4DNA末端的单链DNA(ssDNA)；且1个解旋酶核心单体可与G4DNA3'或5'末端10个核苷酸的ssDNA结合。BLM解旋酶催化核心对G4DNA的解链依赖于3'-末端ssDNA和ATP；只具有3'末端ssDNA的G4DNA是解旋酶解链的最理想底物，且有效解链需要的3'末端ssDNA的长度为10 个核苷酸。解旋酶也能够与钝末端的G4DNA松弛地结合且对其部分解链。这些结果对于进一步研究BLM解旋酶在细胞中的功能具有一定帮助。

关键词：结合 BLM解旋酶 G4DNA 解链荧光偏振

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LUO Heng,XU Houqiang,CAI Mingjuan,et al. The binding and unwinding properties of the Bloom helicase catalytic core to the G4DNA structure[EB/OL]. Beijing:Sciencepaper Online[2012-03-22]. https://www.paper.edu.cn/releasepaper/content/201203-631.

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论文编号	201203-631
论文题目	BLM解旋酶催化核心与G4DNA结合和解链特性
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