The Mre11 Nuclease Is Not Required for 5′ to 3′ Resection at Multiple HO-Induced Double-Strand Breaks

Bertrand Llorente; Lorraine S. Symington

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The Mre11 Nuclease Is Not Required for 5′ to 3′ Resection at Multiple HO-Induced Double-Strand Breaks

机译：在多个HO诱导的双链断裂处进行5'至3'切除不需要Mre11核酸酶

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Current hypotheses suggest the Mre11 nuclease activity could be directly involved in double-strand break (DSB) resection in the presence of a large number of DSBs or limited to processing abnormal DNA ends. To distinguish between these possibilities, we used two methods to create large numbers of DSBs in Saccharomyces cerevisiae chromosomes, without introducing other substrates for the Mre11 nuclease. Multiple DSBs were created either by expressing the HO endonuclease in strains containing several HO cut sites embedded within randomly dispersed Ty1 elements or by phleomycin treatment. Analysis of resection by single-strand DNA formation in these systems showed no difference between strains containing MRE11 or the mre11-D56N nuclease defective allele, suggesting that the Mre11 nuclease is not involved in the extensive 5′ to 3′ resection of DSBs. We postulate that the ionizing radiation (IR) sensitivity of mre11 nuclease-defective mutants results from the accumulation of IR-induced DNA damage that is normally processed by the Mre11 nuclease. We also report that the processivity of 5′ to 3′ DSB resection and the yield of repaired products are affected by the number of DSBs in a dose-dependent manner. Finally, we show that the exonuclease Exo1 is involved in the processivity of 5′ to 3′ resection of an HO-induced DSB at the MAT locus.

机译：当前的假设表明，在存在大量DSB或仅限于处理异常DNA末端的情况下，Mre11核酸酶活性可能直接参与双链断裂（DSB）切除。为了区分这些可能性，我们使用了两种方法在啤酒酵母（Saccharomyces cerevisiae）染色体中创建大量DSB，而没有为Mre11核酸酶引入其他底物。通过在包含嵌入随机分散的Ty1元件中的几个HO切割位点的菌株中表达HO核酸内切酶或通过霉素处理来创建多个DSB。在这些系统中通过单链DNA形成进行的切除分析显示，包含 MRE11 或 mre11-D56N 核酸酶缺陷等位基因的菌株之间没有差异，表明Mre11核酸酶不参与DSB的5'至3'广泛切除术中。我们推测 mre11 核酸酶缺陷型突变体的电离辐射（IR）敏感性是由IR诱导的DNA损伤的积累而引起的，该损伤通常由Mre11核酸酶处理。我们还报告说5'至3'DSB切除的进行性和修复产品的产量以剂量依赖的方式受到DSB数量的影响。最后，我们表明核酸外切酶Exo1参与了HO诱导的DSB在 MAT 位点的5'至3'切除的过程。 展开▼

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《Molecular and Cellular Biology》 |2004年第21期|共13页

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Bertrand Llorente; Lorraine S. Symington;
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中图分类细胞生物学;

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6. The Mre11 Nuclease Is Not Required for 5′ to 3′ Resection at Multiple HO-Induced Double-Strand Breaks [O] . Bertrand Llorente, Lorraine S. Symington 2004

机译：在多个HO诱导的双链断裂处进行5至3切除不需要Mre11核酸酶

7. The Mre11 Nuclease Is Not Required for 5′ to 3′ Resection at Multiple HO-Induced Double-Strand Breaks [O] . Llorente, Bertrand, Symington, Lorraine S. 2004

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The Mre11 Nuclease Is Not Required for 5′ to 3′ Resection at Multiple HO-Induced Double-Strand Breaks

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