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Rec2 Interplay with both Brh2 and Rad51 Balances Recombinational Repair in Ustilago maydis

机译:Rec2与Brh2和Rad51的平衡相互作用在乌斯迪亚戈(Ustilago maydis)中实现重组

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摘要

Rec2 is the single Rad51 paralog in Ustilago maydis. Here, we find that Rec2 is required for radiation-induced Rad51 nuclear focus formation but that Rec2 foci form independently of Rad51 and Brh2. Brh2 foci also form in the absence of Rad51 and Rec2. By coprecipitation from cleared extracts prepared from Escherichia coli cells expressing the proteins, we found that Rec2 interacts physically not only with Rad51 and itself but also with Brh2. Transgenic expression of Brh2 in rec2 mutants can effectively restore radiation resistance, but the frequencies of spontaneous Rad51 focus formation and allelic recombination are elevated. The Dss1-independent Brh2-RPA70 fusion protein is also active in restoring radiation sensitivity of rec2 but is hyperactive to an extreme degree in allelic recombination and in suppressing the meiotic block of rec2. However, the high frequency of chromosome missegregation in meiotic products is an indicator of a corrupted process. The results demonstrate that the importance of Rec2 function is not only in stimulating recombination activity but also in ensuring that recombination is properly controlled.
机译:Rec2是Ustilago maydis中的单个Rad51旁系同源物。在这里,我们发现Rec2是辐射诱导的Rad51核焦点形成所必需的,但是Rec2焦点的形成独立于Rad51和Brh2。在没有Rad51和Rec2的情况下也会形成Brh2病灶。通过从表达蛋白质的大肠杆菌细胞制备的澄清提取物中共沉淀,我们发现Rec2不仅与Rad51及其自身发生物理相互作用,而且与Brh2发生物理相互作用。 Brh2在rec2突变体中的转基因表达可以有效地恢复抗辐射性,但自发的Rad51焦点形成和等位基因重组的频率增加。不依赖于Dss1的Brh2-RPA70融合蛋白在恢复rec2的放射敏感性方面也具有活性,但是在等位基因重组和抑制rec2的减数分裂中具有极高的活性。但是,减数分裂产物中染色体错位的高频率是破坏过程的指示。结果表明,Rec2功能的重要性不仅在于刺激重组活性,而且在于确保正确控制重组。

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