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Activation of the ATPase Activity of Adeno-Associated Virus Rep68 and Rep78

机译：Adeno相关病毒REP68和REP78的ATP酶活性的激活

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摘要

Rep68 and Rep78 DNA helicases, encoded by adeno-associated virus 2 (AAV2), are required for replication of AAV viral DNA in infected cells. They bind to imperfect palindromic elements in the inverted terminal repeat structures at the 3'- and 5'-ends of virion DNA. The ATPase activity of Rep68 and Rep78 is stimulated up to 10-fold by DNA containing the target sequence derived from the inverted terminal repeat; nontarget DNA stimulates ATPase activity at 50-fold higher concentrations. Activation of ATPase activity of Rep68 by DNA is cooperative with a Hill coefficient of 1.8 ± 0.2. When examined by gel filtration at 0.5 M NaCl in the absence of DNA, Rep68 self-associates in a concentration-dependent manner. In the presence of DNA containing the binding element, Rep68 (and Rep78) forms protein- DNA complexes that exhibit concentration-dependent self-association in gel filtration analysis. The ATPase activity of the isolated Rep68-DNA and Rep78-DNA complexes is not activated by additional target DNA. Results of sedimentation velocity experiments in the presence of saturating target DNA are consistent with Rep68 forming a hexamer of the protein with two copies of the DNA element. Activation of the ATPase activity of Rep68 is associated with the formation of a protein-DNA oligomer.
机译：通过腺相关病毒2（AAV2）编码的REP68和REP78 DNA螺旋酶是在感染细胞中复制AAV病毒DNA所必需的。它们在Virion DNA的3'-和5'-末端的倒置末端重复结构中结合不完美的回复末端元素。通过含有衍生自倒端子重复的靶序列的DNA刺激REP68和REP78的ATP酶活性高达10倍; Nontarget DNA以50倍的浓度刺激ATP酶活性。 DNA的ATP酶活性的激活是具有1.8±0.2的山坡系数的合作。当在不存在DNA的情况下通过在0.5M NaCl的凝胶过滤时检查，Rep68以浓度依赖性方式自相同。在含有结合元素的DNA存在下，REP68（和REP78）形成蛋白质DNA复合物，其在凝胶过滤分析中表现出浓度依赖性的自我关联。分离的REP68-DNA和REP78-DNA复合物的ATP酶活性未被另外的靶DNA激活。饱和靶DNA存在下沉淀速度实验的结果与克REP68一致地形成蛋白质的六聚体，其中DNA元素的两份。 REP68的ATP酶活性的激活与形成蛋白质-DNA低聚物的形成有关。

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  • 来源
    《Biochemistry》 |2007年第21期|共11页
  • 作者

    Susan S. Dignam; John J. Correia; Shadia E. Nada;

  • 作者单位

    Department of Biochemistry and Cancer Biology University of Toledo College of Medicine 3035 Arlington Avenue Toledo Ohio 43614-5804 and Department of Biochemistry University of Mississippi Medical Center 2500 North State Street Jackson Missi;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物化学;
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