Investigating Deinococcus radiodurans RecA protein filament formation on double-stranded DNA by a real-time single-molecule approach.
Investigating Deinococcus radiodurans RecA protein filament formation on double-stranded DNA by a real-time single-molecule approach.
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DOI：
10.1021/bi200423t
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发表时间：
2011-10-04
期刊：
影响因子：
2.9
通讯作者：
Li, Hung-Wen
中科院分区：
文献类型：
--
作者：
Hsu, Hsin-Fang;Ngo, Khanh V.;Chitteni-Pattu, Sindhu;Cox, Michael M.;Li, Hung-Wen
在高效、精确且几乎无错误的DNA修复系统的帮助下，耐辐射奇球菌可以在高剂量辐射或干燥造成的数百次双链断裂中存活下来。耐辐射球菌的RecA（DrRecA）在修复的早期阶段通过扩展的合成依赖性链退火过程和在后期更一般的同源重组阶段都起着核心作用。这两种作用可能都需要在双链DNA上形成DrRecA细丝。我们已经开发了单分子拴系粒子运动（TPM）实验，通过观察RecA结合导致的DNA链长的变化来研究RecA蛋白在单个双链DNA分子上的组装动力学。我们证明了DrRecA在dsDNA上的成核比大肠杆菌（Ec）RecA蛋白快得多，但延伸较慢。相对于EcRecA，这种属性的组合往往会增加DrRecA细丝的数量并减少其长度，这一特征可能反映了在辐照的异常球菌细胞中同时修复数百个基因组双链断裂的要求。
With the aid of an efficient, precise, and almost error-free DNA repair system, Deinococcus radiodurans can survive hundreds of double strand breaks inflicted by high doses of irradiation or desiccation. The RecA of Deinococcus radiodurans (DrRecA) plays a central role both in the early phase of repair by an extended synthesis-dependent strand annealing process and in the later more general homologous recombination phase. Both roles likely require DrRecA filament formation on duplex DNA. We have developed single-molecule tethered particle motion (TPM) experiments to study the assembly dynamics of RecA proteins on individual duplex DNA molecules by observing changes in DNA tether length resulting from RecA binding. We demonstrate that DrRecA nucleation on dsDNA is much faster than Escherichia coli (Ec) RecA protein, but the extension is slower. This combination of attributes would tend to increase the number and decrease the length of DrRecA filaments relative to those of EcRecA, a feature that may reflect the requirement to repair hundreds of genomic double strand breaks concurrently in irradiated Deinococcus cells.
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影响因子：
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