Recombination hotspot activity of hypervariable minisatellite DNA requires minisatellite DNA binding proteins.
Recombination hotspot activity of hypervariable minisatellite DNA requires minisatellite DNA binding proteins.
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高变小卫星 DNA 的重组热点活性需要小卫星 DNA 结合蛋白。
DOI：
10.1007/bf02677494
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Moore,PD
中科院分区：
文献类型：
--
作者：
Wahls,WP;Moore,PD
高变小卫星DNA重复序列存在于哺乳动物基因组中的数万个位点。这些序列刺激哺乳动物细胞中的同源重组[Cell 60：95-103]。为了验证蛋白质-DNA相互作用是体内热点功能所必需的这一假设，我们确定了附近的第二个蛋白质结合是否可以消除热点活性。在存在或不存在膜作用重组热点以及存在或不存在第二反式作用DNA结合蛋白的情况下测量质粒底物对之间的分子间重组。小卫星DNA在两个细胞系中具有热点活性，但在两个表达与热点相邻的DNA结合的位点特异性解旋酶的密切相关的细胞系中缺乏热点活性。热点功能的抑制发生复制和非复制重组基板。这些结果表明，体内热点活动需要小卫星DNA结合蛋白占据位点。
Hypervariable minisatellite DNA repeats are found at tens of thousands of loci in the mammalian genome. These sequences stimulate homologous recombination in mammalian cells [Cell60:95–103]. To test the hypothesis that protein-DNA interaction is required for hotspot functionin vivo, we determined whether a second protein binding nearby could abolish hotspot activity. Intermolecular recombination between pairs of plasmid substrates was measured in the presence or absence of thecis-acting recombination hotspot and in the presence or absence of the secondtrans-acting DNA binding protein. Minisatellite DNA had hotspot activity in two cell lines, but lacked hotspot activity in two closely related cell lines expressing a site-specific helicase that bound to DNA adjacent to the hotspot. Suppression of hotspot function occurred for both replicating and non-replicating recombination substrates. These results indicate that hotspot activityin vivorequires site occupancy by minisatellite DNA binding proteins.