BRCA2-DSS1 interaction is dispensable for RAD51 recruitment at replication-induced and meiotic DNA double strand breaks.
BRCA2-DSS1 interaction is dispensable for RAD51 recruitment at replication-induced and meiotic DNA double strand breaks.
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DOI：
10.1038/s41467-022-29409-y
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发表时间：
2022-04-01
影响因子：
16.6
通讯作者：
Sharan SK
中科院分区：
文献类型：
--
作者：
Mishra AP;Hartford SA;Sahu S;Klarmann K;Chittela RK;Biswas K;Jeon AB;Martin BK;Burkett S;Southon E;Reid S;Albaugh ME;Karim B;Tessarollo L;Keller JR;Sharan SK
肿瘤抑制因子 BRCA2 和 DSS1 之间的相互作用对于 RAD51 募集和通过同源重组 (HR) 修复 DNA 双链断裂 (DSB) 至关重要。我们已经培育出 BRCA2 的 2431 位亮氨酸被脯氨酸取代的小鼠，这破坏了这种相互作用。尽管大量突变小鼠在胚胎发生过程中死亡，但一些纯合和半合突变小鼠经历了正常的出生后发育。尽管缺乏辐射诱导的 RAD51 灶形成以及体细胞中严重的 HR 缺陷，但突变小鼠具有生育能力，并且在减数分裂期间表现出正常的 RAD51 募集。我们假设早期 I 期同源染色体的存在可以弥补 BRCA2-DSS1 相互作用的缺陷。我们展示了当拓扑异构酶 I 抑制剂诱导的单链断裂在 DNA 复制过程中转化为 DSB 时，突变细胞中 RAD51 焦点的恢复。我们还通过使用链霉亲和素融合的 Cas9 将供体 DNA 连接到 DSB 位点来部分挽救 HR 缺陷。我们的研究结果表明，当同源 DNA 靠近 DSB 时，BRCA2-DSS1 复合物对于 RAD51 加载来说是可有可无的。米什拉等人。已经培育出 BRCA2 中单个氨基酸取代的小鼠，这破坏了其与 DSS1 的相互作用，导致严重的 HR 缺陷。他们表明，在复制诱导和减数分裂 DSB 中，相互作用对于 HR 来说是可有可无的。
The interaction between tumor suppressor BRCA2 and DSS1 is essential for RAD51 recruitment and repair of DNA double stand breaks (DSBs) by homologous recombination (HR). We have generated mice with a leucine to proline substitution at position 2431 of BRCA2, which disrupts this interaction. Although a significant number of mutant mice die during embryogenesis, some homozygous and hemizygous mutant mice undergo normal postnatal development. Despite lack of radiation induced RAD51 foci formation and a severe HR defect in somatic cells, mutant mice are fertile and exhibit normal RAD51 recruitment during meiosis. We hypothesize that the presence of homologous chromosomes in close proximity during early prophase I may compensate for the defect in BRCA2-DSS1 interaction. We show the restoration of RAD51 foci in mutant cells when Topoisomerase I inhibitor-induced single strand breaks are converted into DSBs during DNA replication. We also partially rescue the HR defect by tethering the donor DNA to the site of DSBs using streptavidin-fused Cas9. Our findings demonstrate that the BRCA2-DSS1 complex is dispensable for RAD51 loading when the homologous DNA is close to the DSB. Mishra et al. have generated mice with a single amino acid substitution in BRCA2, which disrupts its interaction with DSS1 resulting in a severe HR defect. They show the interaction to be dispensable for HR at replication induced and meiotic DSBs.
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影响因子：
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