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Atomic force microscopic measurement of the interdomain angle in symmetric holliday junctions

机译:对称Holliday联系中的互联角度的原子力显微镜测量

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摘要

The Holliday junction is a key intermediate in genetic recombination.It consists of four DNA strands that associate by base pairing to produce four double helices flanking a junction point.In the presence of multivalent cations,the four helices,in turn,stack in pairs to formtwo double-helical domains.The angle between these domains has been shown in a number of solution studies to be approx60deg in junctions flanked by asymmetric sequences.However,the recently determined crystal structure of a symmetric junction finds an angle closer to 40deg,possiblybecause of sequence effects.From the crystal structure alone,one cannot exclude the possibility that this unusual angle is a consequence of crystal packing effects.We have formed two-dimensional (2D)periodic arrays of DNA parallelograms with the same junction-flanking sequence used to produce thecrystals;these parallelograms are free to adopt their preferred interdomain angle.Atomic force microscopy can be used to establishe the interdomain angle in this system.We find that the angle in this junction is 43deg,in good agreement with the results of crystallography.We have used hydroxyl radical autofootprinting to establish that the branchpoint is at the same migratory position seen in the crystals.
机译:Holliday结是遗传重组的关键中间体。它由四个DNA股线组成,其通过基部配对赋予侧翼的四个双螺旋。在多价阳离子的存在下,四个螺旋,又成对堆叠formtwo双螺旋域。这些域之间的角度已经显示在多个解决方案研究中,在非对称序列中侧翼的结侧的几个解决方案研究中。然而,最近确定的对称结的晶体结构发现更靠近40deg的角度,可能是序列效果。通过单独的晶体结构,一个不能排除这种异常角度是晶体填料效果的结果的可能性。我们已经形成二维(2D)与用于生产相同的结侧序列的DNA平行四边形阵列形成TheCryStals;这些平行四边形是自由的,采用优选的跨域角度。atomic力显微镜可用于建立Interbada N角度在该系统中。我们发现该结的角度是43deg,与晶体学结果吻合良好。我们使用羟基自由基自身吸引力,以确定分支位于晶体中看到的相同迁移位置。

著录项

  • 来源
    《Biochemistry》 |2002年第19期|共6页
  • 作者单位

    Department of Chemistry New York University New York New York 10003;

    Department of Chemistry New York University New York New York 10003;

    Department of Chemistry New York University New York New York 10003;

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

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