The Mre11 protein interacts with both Rad50 and the HerA bipolar helicase and is recruited to DNA following gamma irradiation in the archaeon Sulfolobus acidocaldarius.
The Mre11 protein interacts with both Rad50 and the HerA bipolar helicase and is recruited to DNA following gamma irradiation in the archaeon Sulfolobus acidocaldarius.
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DOI：
10.1186/1471-2199-9-25
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发表时间：
2008-02-22
影响因子：
--
通讯作者：
Elie C
中科院分区：
文献类型：
--
作者：
Quaiser A;Constantinesco F;White MF;Forterre P;Elie C
普遍存在的Rad 50和Mre 11蛋白在细菌和真核生物中维持基因组完整性的许多过程中发挥关键作用，但它们在真核生物中的功能目前尚不清楚。我们先前表明，在大多数极端嗜热古菌中，rad 50-mre 11基因与编码单链核酸内切酶和5'至3'核酸外切酶的nurA和编码双极DNA解旋酶的herA相连，这表明这四种蛋白质参与了共同的分子途径。由于超嗜热古菌的遗传工具刚刚出现，我们利用免疫检测方法来获得这些蛋白在超嗜热泉古菌酸热硫化叶菌中的作用的第一个体内数据。我们首先证明了S. acidocaldarius可以修复高剂量γ射线诱导的DNA损伤，我们进行了一个时间过程分析的总水平和亚细胞分配的Rad 50，Mre 11，HerA和NurA沿着与RadA重组酶在控制和照射细胞。我们发现，在指数阶段，所有的蛋白质合成和显示恒定的水平，但它们都表现出不同的亚细胞分配。γ射线照射后，Mre 11和RadA都立即被募集到DNA中，并在DNA修复过程中保持DNA结合。此外，我们通过免疫沉淀试验表明，Rad 50，Mre 11和HerA解旋酶相互作用。我们的分析强烈支持，在酸热硫化叶菌，Mre 11蛋白和RadA重组酶可能发挥积极的作用，在修复由γ射线和/或可能作为DNA损伤传感器引入的DNA损伤。此外，我们的研究结果表明Mre 11，Rad 50和HerA解旋酶之间的功能相互作用，并表明每个蛋白质发挥不同的作用时，其本身或与其合作伙伴。本报告提供了第一个在体内的证据支持的含义Mre 11蛋白在DNA修复过程中的dendritea和显示其相互作用与Rad 50和HerA双极解旋酶。进一步研究这些蛋白质，NurA核酸酶和RadA重组酶之间的功能相互作用，将使我们能够定义它们的作用和作用机制。
The ubiquitous Rad50 and Mre11 proteins play a key role in many processes involved in the maintenance of genome integrity in Bacteria and Eucarya, but their function in the Archaea is presently unknown. We showed previously that in most hyperthermophilic archaea, rad50-mre11 genes are linked to nurA encoding both a single-strand endonuclease and a 5' to 3' exonuclease, and herA, encoding a bipolar DNA helicase which suggests the involvement of the four proteins in common molecular pathway(s). Since genetic tools for hyperthermophilic archaea are just emerging, we utilized immuno-detection approaches to get the first in vivo data on the role(s) of these proteins in the hyperthermophilic crenarchaeon Sulfolobus acidocaldarius. We first showed that S. acidocaldarius can repair DNA damage induced by high doses of gamma rays, and we performed a time course analysis of the total levels and sub-cellular partitioning of Rad50, Mre11, HerA and NurA along with the RadA recombinase in both control and irradiated cells. We found that during the exponential phase, all proteins are synthesized and display constant levels, but that all of them exhibit a different sub-cellular partitioning. Following gamma irradiation, both Mre11 and RadA are immediately recruited to DNA and remain DNA-bound in the course of DNA repair. Furthermore, we show by immuno-precipitation assays that Rad50, Mre11 and the HerA helicase interact altogether. Our analyses strongly support that in Sulfolobus acidocaldarius, the Mre11 protein and the RadA recombinase might play an active role in the repair of DNA damage introduced by gamma rays and/or may act as DNA damage sensors. Moreover, our results demonstrate the functional interaction between Mre11, Rad50 and the HerA helicase and suggest that each protein play different roles when acting on its own or in association with its partners. This report provides the first in vivo evidence supporting the implication of the Mre11 protein in DNA repair processes in the Archaea and showing its interaction with both Rad50 and the HerA bipolar helicase. Further studies on the functional interactions between these proteins, the NurA nuclease and the RadA recombinase, will allow us to define their roles and mechanism of action.
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