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国家自然科学基金(30700011)

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发文基金:国家自然科学基金国家重点基础研究发展计划更多>>
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Co-expression with RadA and the characterization of stRad55B, a RadA paralog from the hyperthermophilic crenarchaea Sulfolobus tokodaii
2008年
ST0838 (designed stRad55B) is one of the four RadA paralogs (or Rad55 homologues) in the genome of the hyperthermophilic crenarchaeon Sulfolobus tokodaii. The gene is induced by UV irradiation, sug-gesting that it is involved in DNA recombinational repair in this organism. However, this protein could not be expressed normally in vitro. In this study, thermostable and soluble stRad55B was obtained by co-expression with S. tokodaii RadA (stRadA) in E. coli, and the enzymatic properties were examined. It was found that stRad55B bound ssDNA preferentially and had a very weak ATPase activity that was not stimulated by DNA. The recombinant protein inhibited the strand exchange activity promoted by stRadA, indicating that stRad55B might be an inhibitor to the homologous recombination in this ar-chaeon. The results will be helpful for further functional and interaction analysis of RadA paralogs and for the understanding of the mechanism of recombinational repair in archaea.
SHENG DuoHong, LI MingFeng, JIAO JianDong, NI JinFeng & SHEN YuLong State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China
关键词:ARCHAEAREPAIRRADACO-EXPRESSION
泉古菌Sulfolobus tokodaii RadA同系物stRad55B与RadA的共表达及其对RadA重组活性的抑制
2008年
ST0838(定义为stRad55B)是超嗜热古菌(Sulfolobus tokodaii)编码的4个RadA的同系物(或Rad55同源蛋白)之一.研究发现,它能够被紫外线(UV)辐射损伤诱导,可能参与了细胞内的DNA损伤修复过程,然而利用常规方法,该蛋白不能体外可溶性地表达.通过和RadA共表达,得到了具有热稳定性的可溶stRad55B蛋白,并对其活性进行了初步检测.stRad55B优先结合单链DNA,并且具有不依赖DNA的ATP酶活性.另外,DNA链交换实验发现stRad55B能够明显抑制RadA催化的链重组活性,表现出一个重组修复系统抑制蛋白的特征.实验结果为进一步研究古菌中RadA同系蛋白的功能以及相互作用机制,揭示古菌DNA同源重组修复机理提供了依据.
盛多红李铭峰焦建东倪金凤申玉龙
关键词:古菌RADA共表达
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