Exo1-protected DNA nicks direct crossover formation in meiosis

Exo1-protected DNA nicks direct crossover formation in meiosis
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Exo1 保护的 DNA 切口在减数分裂中直接交叉形成

DOI:
10.1101/2021.08.29.458102
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发表时间:
2021
期刊:
bioRxiv
影响因子:
--
通讯作者:
Gioia M, Payero L
Gioia M, Payero L
中科院分区:
--
文献类型:
--
作者:
Gioia M, Payero L

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在大多数有性生殖的生物中,减数分裂期间同源染色体之间的交换是产生单倍体配子所必需的。在芽殖酵母减数分裂中形成的大多数交换是由双霍利迪连接(dHJ)中间体的偏向性解析引起的。该dHJ解析步骤涉及Rad 2/XPG家族核酸酶Exo 1和Mlh 1-Mlh 3错配修复核酸内切酶的作用。在这里,我们提供了面包酵母中的遗传证据,证明Exo 1通过保护DNA切口免受连接来促进减数分裂交换。我们发现,与DNA相互作用的Exo 1中的结构元件,例如在切口/皮瓣识别过程中弯曲DNA所需的结构元件,对于其在交叉中的作用至关重要。与这些观察结果相一致,Rad 2/XPG家族成员Rad 27的减数分裂表达部分挽救了exo 1 null突变体中的交叉缺陷,Cdc 9连接酶的减数分裂过表达降低了exo 1 DNA结合突变体的交叉水平,使其接近exo 1 null。此外,我们的工作确定了exo 1在交叉干扰中的作用。总之,这些研究为Exo 1保护的切口对于减数分裂交换的形成及其分布至关重要提供了实验证据。
In most sexually reproducing organisms crossing over between chromosome homologs during meiosis is essential to produce haploid gametes. Most crossovers that form in meiosis in budding yeast result from the biased resolution of double Holliday junction (dHJ) intermediates. This dHJ resolution step involves the actions of Rad2/XPG family nuclease Exo1 and the Mlh1-Mlh3 mismatch repair endonuclease. Here, we provide genetic evidence in baker’s yeast that Exo1 promotes meiotic crossing over by protecting DNA nicks from ligation. We found that structural elements in Exo1 that interact with DNA, such as those required for the bending of DNA during nick/flap recognition, are critical for its role in crossing over. Consistent with these observations, meiotic expression of the Rad2/XPG family member Rad27 partially rescued the crossover defect inexo1null mutants, and meiotic overexpression of Cdc9 ligase reduced the crossover levels ofexo1DNA-binding mutants to levels that approached theexo1null. In addition, our work identified a role for Exo1 in crossover interference. Together, these studies provide experimental evidence for Exo1-protected nicks being critical for the formation of meiotic crossovers and their distribution.
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