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首页> 外文期刊>Biochemistry >The Deinococcus radiodurans-Encoded HU Protein Has Two DNA-Binding Domains
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The Deinococcus radiodurans-Encoded HU Protein Has Two DNA-Binding Domains

机译：Deinococcus radiodurans编码的HU蛋白具有两个DNA结合域

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摘要

Deinococcus radiodurans can reconstitute its genome from double-strand breaks,most likely due to unusually efficient DNA repair and recombination.Factors that may contribute to such processes include the histone-like protein HU.The D.radiodurans-encoded HU (DrHU),which binds preferentially to DNA recombination intermediates,contains a 47-amino acid extension preceding the fold characteristic of HU proteins.Here we use electrophoretic mobility shift assays and DNA footprinting to show that the DrHU N-terminal domain significantly modulates DNA binding.The truncated DrHU (DELTADrHU),comprising only the conserved DNA-binding fold,has a site size of approx 11 bp in contrast to full-length DrHU which does not stably engage DNA shorter than approx 50 bp.Unlike wild-type DrHU,ADrHU distinguishes between linear DNA and DNA with nicks or gaps.ADrHU also binds preferentially to four-way junction (4WJ) DNA,with half-maximal saturation of 1.4 +- 0.4 nM compared to 20 +- 2 nM for 37 bp duplex DNA.However,in contrast to full-length protein which binds the junction arms,ADrHU primarily protects the junction crossover.Evidently,the DrHU N-terminus changes the mode of binding to both 4WJ DNA,duplex DNA,and DNA with nicks or gaps,thereby resulting in DrHU binding preferentially only to 4WJ structures.Combined with Western blots that confirm the presence of the extended form of DrHU in vivo,our data provide mechanistic insight into discrimination between 4WJ DNA and other distorted DNA constructs and suggest that an in vivo role of DrHU may be to stabilize DNA junctions.
机译：放射球菌可以通过双链断裂来重组其基因组，这很可能是由于异常有效的DNA修复和重组所致。可能导致这种过程的因素包括组蛋白样蛋白质HU.D.radiodurans编码的HU（DrHU），优先与DNA重组中间体结合，在HU蛋白的折叠特征之前包含47个氨基酸的延伸。在这里，我们使用电泳迁移率变动分析和DNA足迹检测显示DrHU N末端结构域显着调节DNA结合。 DELTADrHU）仅包含保守的DNA结合折叠，其位点大小约为11 bp，而全长DrHU不能稳定地结合短于约50 bp的DNA。与野生型DrHU不同，ADrHU区分线性DNA ADrHU还优先结合四向连接（4WJ）DNA，其最大半饱和度为1.4±0.4 nM，而37 bp双链DNA的最大饱和度为20±2 nM。但是，与结合连接臂的全长蛋白相反，ADrHU主要保护连接交叉。显然，DrHU N末端会改变与4WJ DNA，双链DNA以及带有缺口或缺口的DNA的结合方式，从而因此，我们的数据为区分4WJ DNA和其他扭曲的DNA构建体提供了机制上的洞察力，并暗示了DrHU在体内的作用。 DrHU可能是为了稳定DNA连接。

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