{"title":"超变异小卫星DNA的重组热点活性需要小卫星DNA结合蛋白。","authors":"W P Wahls, P D Moore","doi":"10.1007/BF02677494","DOIUrl":null,"url":null,"abstract":"<p><p>Hypervariable minisatellite DNA repeats are found at tens of thousands of loci in the mammalian genome. These sequences stimulate homologous recombination in mammalian cells [Cell 60:95-103]. To test the hypothesis that protein-DNA interaction is required for hotspot function in 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 the cis-acting recombination hotspot and in the presence or absence of the second trans-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 activity in vivo requires site occupancy by minisatellite DNA binding proteins.</p>","PeriodicalId":21884,"journal":{"name":"Somatic Cell and Molecular Genetics","volume":"24 1","pages":"41-51"},"PeriodicalIF":0.0000,"publicationDate":"1998-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3151739/pdf/nihms311591.pdf","citationCount":"0","resultStr":"{\"title\":\"Recombination hotspot activity of hypervariable minisatellite DNA requires minisatellite DNA binding proteins.\",\"authors\":\"W P Wahls, P D Moore\",\"doi\":\"10.1007/BF02677494\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Hypervariable minisatellite DNA repeats are found at tens of thousands of loci in the mammalian genome. These sequences stimulate homologous recombination in mammalian cells [Cell 60:95-103]. To test the hypothesis that protein-DNA interaction is required for hotspot function in 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 the cis-acting recombination hotspot and in the presence or absence of the second trans-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 activity in vivo requires site occupancy by minisatellite DNA binding proteins.</p>\",\"PeriodicalId\":21884,\"journal\":{\"name\":\"Somatic Cell and Molecular Genetics\",\"volume\":\"24 1\",\"pages\":\"41-51\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1998-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3151739/pdf/nihms311591.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Somatic Cell and Molecular Genetics\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1007/BF02677494\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Somatic Cell and Molecular Genetics","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/BF02677494","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
哺乳动物基因组中有数以万计的位点存在超变异小卫星 DNA 重复序列。这些序列刺激哺乳动物细胞中的同源重组 [细胞 60:95-103]。为了验证体内热点功能需要蛋白质-DNA 相互作用的假设,我们测定了附近的第二种蛋白质结合是否会削弱热点的活性。在存在或不存在顺式作用重组热点以及存在或不存在第二种反式作用 DNA 结合蛋白的情况下,测量了成对质粒底物之间的分子间重组。小卫星 DNA 在两种细胞系中具有热点活性,但在两种表达与热点邻近 DNA 结合的位点特异性螺旋酶的密切相关细胞系中缺乏热点活性。复制和非复制重组底物都会抑制热点功能。这些结果表明,体内的热点活性需要小卫星DNA结合蛋白的位点占据。
Recombination hotspot activity of hypervariable minisatellite DNA requires minisatellite DNA binding proteins.
Hypervariable minisatellite DNA repeats are found at tens of thousands of loci in the mammalian genome. These sequences stimulate homologous recombination in mammalian cells [Cell 60:95-103]. To test the hypothesis that protein-DNA interaction is required for hotspot function in 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 the cis-acting recombination hotspot and in the presence or absence of the second trans-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 activity in vivo requires site occupancy by minisatellite DNA binding proteins.