Numerical and spatial patterning of yeast meiotic DNA breaks by Tel1.
Numerical and spatial patterning of yeast meiotic DNA breaks by Tel1.
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DOI：
10.1101/gr.213587.116
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发表时间：
2017-02
期刊：
影响因子：
7
通讯作者：
Keeney S
中科院分区：
文献类型：
--
作者：
Mohibullah N;Keeney S
Spo 11产生的启动减数分裂重组的双链断裂（DSB）是危险的病变，可以破坏基因组的完整性，因此减数分裂细胞调节它们的数量，时间和分布。这种调节的机制仍然知之甚少。在这里，我们使用Spo 11-寡核苷酸复合物，DSB形成的副产品，揭示酿酒酵母DNA损伤响应激酶Tel 1（哺乳动物ATM的直系同源物）的贡献方面。一个tel 1 Δ突变体的Spo 11-寡核苷酸复合物的数量总体上增加，并改变了Spo 11-寡核苷酸的长度，这与Tel 1在控制DSB数量和加工中的保守作用一致。激酶死亡的tel 1突变类似地增加Spo 11-寡核苷酸水平，但突变Hop 1和Rec 114上的已知Tel 1磷酸化靶点不增加Spo 11-寡核苷酸水平，这暗示了Tel 1激酶活性并澄清了Tel 1磷酸化底物的作用。Spo 11寡核苷酸的深度测序表明，Tel 1以意想不到的方式塑造了全基因组DSB景观。在减数分裂的早期，Tel 1的缺失引起了大染色体域中DSB分布的广泛变化。然而，随着减数分裂的进行，许多这些变化都会被消除，这说明了DSB调节系统的稳态行为。我们进一步发现，Tel 1的作用是不同的，但与先前描述的重组调节因子Cst 9（也称为Zip 3）的作用部分重叠。最后，我们提供的证据表明，Tel 1依赖的DSB干扰影响人口平均DSB景观，但也表明，人工热点插入的局部抑制作用可以是Tel 1独立和染色体上下文依赖。我们的研究结果描绘Tel 1的作用，在调节的数量和位置的DSB和照明Tel 1和其他途径的DSB控制之间的复杂的相互作用。
The Spo11-generated double-strand breaks (DSBs) that initiate meiotic recombination are dangerous lesions that can disrupt genome integrity, so meiotic cells regulate their number, timing, and distribution. Mechanisms of this regulation remain poorly understood. Here, we use Spo11-oligonucleotide complexes, a byproduct of DSB formation, to reveal aspects of the contribution of the Saccharomyces cerevisiae DNA damage-responsive kinase Tel1 (ortholog of mammalian ATM). A tel1Δ mutant has globally increased amounts of Spo11-oligonucleotide complexes and altered Spo11-oligonucleotide lengths, consistent with conserved roles for Tel1 in control of DSB number and processing. A kinase-dead tel1 mutation similarly increases Spo11-oligonucleotide levels but mutating known Tel1 phosphotargets on Hop1 and Rec114 does not, implicating Tel1 kinase activity and clarifying roles of Tel1 phosphorylation substrates. Deep sequencing of Spo11 oligonucleotides demonstrates that Tel1 shapes the genome-wide DSB landscape in unexpected ways. Early in meiosis, Tel1 absence causes widespread changes in DSB distributions across large chromosomal domains. Many of these changes are erased as meiosis proceeds, however, illustrating homeostatic behavior of DSB regulatory systems. We further find that effects of Tel1 are distinct but partially overlapping with previously described contributions of the recombination regulator Cst9 (also known as Zip3). Finally, we provide evidence indicating that Tel1-dependent DSB interference influences the population-average DSB landscape but also demonstrate that locally inhibitory effects of an artificial hotspot insertion can be both Tel1-independent and chromosomal context-dependent. Our findings delineate Tel1 roles in regulating number and location of DSBs and illuminate the complex interplay between Tel1 and other pathways for DSB control.