Budding yeast ATM/ATR control meiotic double-strand break (DSB) levels by down-regulating Rec114, an essential component of the DSB-machinery.
Budding yeast ATM/ATR control meiotic double-strand break (DSB) levels by down-regulating Rec114, an essential component of the DSB-machinery.
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DOI：
10.1371/journal.pgen.1003545
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发表时间：
2013-06
期刊：
影响因子：
4.5
通讯作者：
Cha RS
中科院分区：
文献类型：
--
作者：
Carballo JA;Panizza S;Serrentino ME;Johnson AL;Geymonat M;Borde V;Klein F;Cha RS
减数分裂的一个基本特征是Spo 11催化程序性DNA双链断裂（DSB）。有证据表明，每次减数分裂产生的DSB的数量是由遗传决定的，并且这种维持预定DSB水平或“DSB稳态”的能力可能是减数分裂程序的性质。在这里，我们提出了直接的证据表明，Rec 114，一个进化上保守的必要组成部分的减数分裂DSB机械，与DSB热点DNA相互作用，Tel 1和Mec 1，芽殖酵母ATM和ATR，分别下调Rec 114减数分裂DSB形成后，通过磷酸化。模拟组成性磷酸化减少了Rec 114和DSB热点DNA之间的相互作用，导致DSB形成的减少和/或延迟。相反，不可磷酸化的rec 114等位基因赋予DSB水平和Rec 114与DSB热点DNA之间的相互作用的全基因组增加。这些观察结果强烈表明，Tel 1和/或Mec 1磷酸化Rec 114后Spo 11催化下调DSB的形成，通过限制Rec 114和DSB热点之间的相互作用。我们还提出的证据表明，Ndt 80，减数分裂特异性转录因子，有助于Rec 114降解，符合其要求完全停止DSB形成。Rec 114病灶从染色质的损失与同源突触，但独立于Ndt 80或Tel 1/Mec 1磷酸化。两者合计，我们提出的证据，三个独立的方式调节Rec 114的活动，这可能有助于减数分裂DSB的稳态，在维持遗传决定的水平的休息。减数分裂是一种支持有性生殖的特殊细胞分裂。它开始于一个二倍体细胞携带两个亲本拷贝的每一条染色体，并结束于四个单倍体细胞，每个细胞只包含一个拷贝。减数分裂的一个基本特征是减数分裂重组，在此过程中，DNA双链断裂（DSB）的程序化产生之后是两个亲本同源物之间的交换，这有助于它们正确分布到子核。不能产生DSB会导致同源分离中的错误，从而产生不能存活的配子。虽然DSB对于减数分裂是必不可少的，但每次断裂都代表着潜在的致命损伤;因此，它的形成必须受到严格的调控。进化上保守的ATM/ATR家族蛋白参与了这种控制;然而，这种控制的机制仍然是难以捉摸的。在这里，我们表明，Tel 1/Mec 1下调减数分裂DSB的形成磷酸化Rec 114，Spo 11复合物的重要组成部分。我们还观察到，Rec 114的活性可以进一步下调，其从染色体中去除，并随后在减数分裂后期的降解。这里提出的证据提供了一个深入了解的方式，其中减数分裂DSB的数量可能会保持在发育编程水平。
An essential feature of meiosis is Spo11 catalysis of programmed DNA double strand breaks (DSBs). Evidence suggests that the number of DSBs generated per meiosis is genetically determined and that this ability to maintain a pre-determined DSB level, or “DSB homeostasis”, might be a property of the meiotic program. Here, we present direct evidence that Rec114, an evolutionarily conserved essential component of the meiotic DSB-machinery, interacts with DSB hotspot DNA, and that Tel1 and Mec1, the budding yeast ATM and ATR, respectively, down-regulate Rec114 upon meiotic DSB formation through phosphorylation. Mimicking constitutive phosphorylation reduces the interaction between Rec114 and DSB hotspot DNA, resulting in a reduction and/or delay in DSB formation. Conversely, a non-phosphorylatable rec114 allele confers a genome-wide increase in both DSB levels and in the interaction between Rec114 and the DSB hotspot DNA. These observations strongly suggest that Tel1 and/or Mec1 phosphorylation of Rec114 following Spo11 catalysis down-regulates DSB formation by limiting the interaction between Rec114 and DSB hotspots. We also present evidence that Ndt80, a meiosis specific transcription factor, contributes to Rec114 degradation, consistent with its requirement for complete cessation of DSB formation. Loss of Rec114 foci from chromatin is associated with homolog synapsis but independent of Ndt80 or Tel1/Mec1 phosphorylation. Taken together, we present evidence for three independent ways of regulating Rec114 activity, which likely contribute to meiotic DSBs-homeostasis in maintaining genetically determined levels of breaks. Meiosis is a specialized cell division that underpins sexual reproduction. It begins with a diploid cell carrying both parental copies of each chromosome, and ends with four haploid cells, each containing only one copy. An essential feature of meiosis is meiotic recombination, during which the programmed generation of DNA double-strand-breaks (DSBs) is followed by the production of crossover(s) between two parental homologs, which facilitates their correct distribution to daughter nuclei. Failure to generate DSBs leads to errors in homolog disjunction, which produces inviable gametes. Although DSBs are essential for meiosis, each break represents a potentially lethal damage; as such, its formation must be tightly regulated. The evolutionarily conserved ATM/ATR family proteins were implicated in this control; nevertheless, the mechanism by which such control could be implemented remains elusive. Here we demonstrate that Tel1/Mec1 down-regulate meiotic DSB formation by phosphorylating Rec114, an essential component of the Spo11 complex. We also observed that Rec114 activity can be further down-regulated by its removal from chromosomes and subsequent degradation during later stages in meiosis. Evidence presented here provides an insight into the ways in which the number of meiotic DSBs might be maintained at developmentally programmed level.
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影响因子：
9.8
作者：
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通讯作者：
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影响因子：
56.9
作者：
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Lichten, M
DOI：
10.1083/jcb.201104121
发表时间：
2011-10-31
期刊：
The Journal of cell biology
影响因子：
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通讯作者：
McKim KS
影响因子：
56.9
作者：
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影响因子：
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通讯作者：
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