The DNA binding properties of the Escherichia coli RecQ helicase
文献类型：期刊论文
| 作者 | Dou, SX ; Wang, PY ; Xu, HQ ; Xi, XG |
| 刊名 | JOURNAL OF BIOLOGICAL CHEMISTRY
 |
| 出版日期 | 2004 |
| 卷号 | 279期号:8页码:6354 |
| 关键词 | SINGLE-STRANDED-DNA WERNER-SYNDROME PROTEIN SYNDROME GENE-PRODUCT NUCLEIC-ACID INTERACTIONS TOPOISOMERASE-III-ALPHA FLUORESCENCE ANISOTROPY EQUILIBRIUM BINDING REP HELICASE ILLEGITIMATE RECOMBINATION REPLICATION CHECKPOINT |
| ISSN号 | 0021-9258 |
| 通讯作者 | Xi, XG (reprint author), Ecole Normale Super, CNRS, UMR 8113, Lab Biotechnol & Pharmacol Genet Appl, 61 Ave President Wilson, F-94235 Cachan, France. |
| 中文摘要 | The RecQ helicase family is highly conserved from bacteria to men and plays a conserved role in the preservation of genome integrity. Its deficiency in human cells leads to a marked genomic instability that is associated with premature aging and cancer. To determine the thermodynamic parameters for the interaction of Escherichia coli RecQ helicase with DNA, equilibrium binding studies have been performed using the thermodynamic rigorous fluorescence titration technique. Steady-state fluorescence anisotropy measurements of fluorescein-labeled oligonucleotides revealed that RecQ helicase bound to DNA with an apparent binding stoichiometry of 1 protein monomer/10 nucleotides. This stoichiometry was not altered in the presence of AMPPNP (adenosine 5'-(beta,gamma-imido) triphosphate) or ADP. Analyses of RecQ helicase interactions with oligonucleotides of different lengths over a wide range of pH, NaCl, and nucleic acid concentrations indicate that the RecQ helicase has a single strong DNA binding site with an association constant at 25 degreesC of K = 6.7 +/- 0.95 x 10(6) M-1 and a cooperativity parameter of omega = 25.5 +/- 1.2. Both single-stranded DNA and double-stranded DNA bind competitively to the same site. The intrinsic affinities are salt-dependent, and the formation of DNA-helicase complex is accompanied by a net release of 3-4 ions. Allosteric effects of nucleotide cofactors on RecQ binding to DNA were observed only for single-stranded DNA in the presence of 1.5 mm AMPPNP, whereas both AMPPNP and ADP had no detectable effect on double-stranded DNA binding over a large range of nucleotide cofactor concentrations. |
| 收录类别 | SCI |
| 语种 | 英语 |
| 公开日期 | 2013-09-23 |
| 源URL | [http://ir.iphy.ac.cn/handle/311004/44998]  |
| 专题 | 物理研究所_物理所公开发表论文_物理所公开发表论文_期刊论文 |
| 推荐引用方式 GB/T 7714 | Dou, SX,Wang, PY,Xu, HQ,et al. The DNA binding properties of the Escherichia coli RecQ helicase[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2004,279(8):6354. |
| APA | Dou, SX,Wang, PY,Xu, HQ,&Xi, XG.(2004).The DNA binding properties of the Escherichia coli RecQ helicase.JOURNAL OF BIOLOGICAL CHEMISTRY,279(8),6354. |
| MLA | Dou, SX,et al."The DNA binding properties of the Escherichia coli RecQ helicase".JOURNAL OF BIOLOGICAL CHEMISTRY 279.8(2004):6354. |
入库方式： OAI收割
来源：物理研究所
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