Ben-Aroya Shay, Mieczkowski Piotr A, Petes Thomas D, Kupiec Martin
Department of Molecular Microbiology and Biotechnology, Tel Aviv University, Ramat Aviv 69978, Israel.
Mol Cell. 2004 Jul 23;15(2):221-31. doi: 10.1016/j.molcel.2004.06.002.
Recombination between repeated DNA sequences can have drastic consequences on the integrity of the genome. Repeated sequences are abundant in most eukaryotes, yet the mechanism that prevents recombination between them is currently unknown. Ty elements, the main family of dispersed repeats in Saccharomyces cerevisiae, exhibit low levels of exchange. Other regions in the genome have relatively high rates of meiotic recombination (hotspots). We show that a Ty element adjacent to the HIS4 recombination hotspot substantially reduces its activity, eliminating local DSB formation. We demonstrate that the Ty has a closed (nuclease-insensitive) chromatin configuration that is also imposed on the flanking DNA sequences. The compact chromatin structure is determined by sequences at the N terminus of the Ty. Increased binding of the Rap1 protein to the hotspot restores both open chromatin conformation and DSB formation. The chromatin configuration of Ty elements precludes initiation of recombination, thus preventing potentially lethal exchanges between repeated sequences.
重复DNA序列之间的重组会对基因组的完整性产生严重影响。重复序列在大多数真核生物中都很丰富,但目前尚不清楚阻止它们之间重组的机制。Ty元件是酿酒酵母中主要的分散重复序列家族,其交换水平较低。基因组中的其他区域具有相对较高的减数分裂重组率(热点)。我们发现,与HIS4重组热点相邻的Ty元件会大幅降低其活性,消除局部双链断裂(DSB)的形成。我们证明,Ty具有封闭(对核酸酶不敏感)的染色质构型,这种构型也施加于侧翼DNA序列。紧密的染色质结构由Ty N端的序列决定。Rap1蛋白与热点的结合增加可恢复开放的染色质构象和DSB的形成。Ty元件的染色质构型可阻止重组的起始,从而防止重复序列之间发生潜在的致死性交换。