Borde V, Wu T C, Lichten M
Laboratory of Biochemistry, Division of Basic Science, National Cancer Institute, Bethesda, Maryland 20892, USA.
Mol Cell Biol. 1999 Jul;19(7):4832-42. doi: 10.1128/MCB.19.7.4832.
In Saccharomyces cerevisiae, meiotic recombination is initiated by DNA double-strand breaks (DSBs). DSBs usually occur in intergenic regions that display nuclease hypersensitivity in digests of chromatin. DSBs are distributed nonuniformly across chromosomes; on chromosome III, DSBs are concentrated in two "hot" regions, one in each chromosome arm. DSBs occur rarely in regions within about 40 kb of each telomere and in an 80-kb region in the center of the chromosome, just to the right of the centromere. We used recombination reporter inserts containing arg4 mutant alleles to show that the "cold" properties of the central DSB-deficient region are imposed on DNA inserted in the region. Cold region inserts display DSB and recombination frequencies that are substantially less than those seen with similar inserts in flanking hot regions. This occurs without apparent change in chromatin structure, as the same pattern and level of DNase I hypersensitivity is seen in chromatin of hot and cold region inserts. These data are consistent with the suggestion that features of higher-order chromosome structure or chromosome dynamics act in a target sequence-independent manner to control where recombination events initiate during meiosis.
在酿酒酵母中,减数分裂重组由DNA双链断裂(DSB)引发。DSB通常发生在染色质消化时显示核酸酶超敏性的基因间区域。DSB在染色体上分布不均;在第三条染色体上,DSB集中在两个“热点”区域,每个染色体臂各有一个。在每条端粒约40 kb范围内的区域以及染色体中心、着丝粒右侧80 kb的区域中,DSB很少发生。我们使用含有arg4突变等位基因的重组报告插入片段来表明,中央DSB缺陷区域的“冷”特性会施加于插入该区域的DNA上。冷区域插入片段显示的DSB和重组频率明显低于侧翼热点区域中类似插入片段的频率。这种情况在染色质结构上没有明显变化,因为在热点和冷区域插入片段的染色质中观察到相同模式和水平的DNase I超敏性。这些数据与以下观点一致,即高阶染色体结构或染色体动力学特征以与靶序列无关的方式起作用,以控制减数分裂期间重组事件起始的位置。