{"title":"Nucleolin 在体外促进同源 DNA 配对。","authors":"B Thyagarajan, R Lundberg, M Rafferty, C Campbell","doi":"10.1023/b:scam.0000007129.98789.1f","DOIUrl":null,"url":null,"abstract":"<p><p>We purified to near homogeneity a previously identified 100 kDa mammalian homologous DNA pairing protein. The purified 100 kDa protein also catalyzed high levels of cell-free homologous DNA recombination activity. This ATP-dependent activity was capable of forming conservative recombinant products between two circular, double-stranded DNA molecules. We were unable to detect any DNA polymerase, DNA ligase, or 5' or 3' exonuclease activity associated with this purified material. The purified 100 kDa protein bound silver nitrate as well as a monoclonal antibody specific for nucleolin. A recombinant protein comprised of the Escherichia coli maltos-ebinding protein fused to the carboxyl-terminal two-thirds of human nucleolin possessed homologous DNA pairing activity. These data indicate that the 100 kDa homologous DNA pairing protein is nucleolin. The observation that nucleolin can carry out homologous DNA strand pairing in vitro raises the prospect that it may function similarly in vivo.</p>","PeriodicalId":21884,"journal":{"name":"Somatic Cell and Molecular Genetics","volume":"24 5","pages":"263-72"},"PeriodicalIF":0.0000,"publicationDate":"1998-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1023/b:scam.0000007129.98789.1f","citationCount":"11","resultStr":"{\"title\":\"Nucleolin promotes homologous DNA pairing in vitro.\",\"authors\":\"B Thyagarajan, R Lundberg, M Rafferty, C Campbell\",\"doi\":\"10.1023/b:scam.0000007129.98789.1f\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>We purified to near homogeneity a previously identified 100 kDa mammalian homologous DNA pairing protein. The purified 100 kDa protein also catalyzed high levels of cell-free homologous DNA recombination activity. This ATP-dependent activity was capable of forming conservative recombinant products between two circular, double-stranded DNA molecules. We were unable to detect any DNA polymerase, DNA ligase, or 5' or 3' exonuclease activity associated with this purified material. The purified 100 kDa protein bound silver nitrate as well as a monoclonal antibody specific for nucleolin. A recombinant protein comprised of the Escherichia coli maltos-ebinding protein fused to the carboxyl-terminal two-thirds of human nucleolin possessed homologous DNA pairing activity. These data indicate that the 100 kDa homologous DNA pairing protein is nucleolin. The observation that nucleolin can carry out homologous DNA strand pairing in vitro raises the prospect that it may function similarly in vivo.</p>\",\"PeriodicalId\":21884,\"journal\":{\"name\":\"Somatic Cell and Molecular Genetics\",\"volume\":\"24 5\",\"pages\":\"263-72\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1998-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1023/b:scam.0000007129.98789.1f\",\"citationCount\":\"11\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Somatic Cell and Molecular Genetics\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1023/b:scam.0000007129.98789.1f\",\"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/10.1023/b:scam.0000007129.98789.1f","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 11
摘要
我们纯化了一种此前已发现的100 kDa哺乳动物同源DNA配对蛋白,其纯度接近均一。纯化的 100 kDa 蛋白还催化了高水平的无细胞同源 DNA 重组活性。这种依赖于 ATP 的活性能够在两个环状双链 DNA 分子之间形成保守的重组产物。我们无法检测到与这种纯化材料相关的任何 DNA 聚合酶、DNA 连接酶或 5' 或 3' 外切酶活性。纯化的 100 kDa 蛋白结合了硝酸银和特异性核仁蛋白单克隆抗体。由大肠杆菌麦芽糖酶结合蛋白与人类核仁蛋白三分之二的羧基末端融合而成的重组蛋白具有同源 DNA 配对活性。这些数据表明,100 kDa 的同源 DNA 配对蛋白就是核仁蛋白。观察到核仁蛋白能在体外进行同源 DNA 链配对,这使人们想到它在体内也可能具有类似的功能。
Nucleolin promotes homologous DNA pairing in vitro.
We purified to near homogeneity a previously identified 100 kDa mammalian homologous DNA pairing protein. The purified 100 kDa protein also catalyzed high levels of cell-free homologous DNA recombination activity. This ATP-dependent activity was capable of forming conservative recombinant products between two circular, double-stranded DNA molecules. We were unable to detect any DNA polymerase, DNA ligase, or 5' or 3' exonuclease activity associated with this purified material. The purified 100 kDa protein bound silver nitrate as well as a monoclonal antibody specific for nucleolin. A recombinant protein comprised of the Escherichia coli maltos-ebinding protein fused to the carboxyl-terminal two-thirds of human nucleolin possessed homologous DNA pairing activity. These data indicate that the 100 kDa homologous DNA pairing protein is nucleolin. The observation that nucleolin can carry out homologous DNA strand pairing in vitro raises the prospect that it may function similarly in vivo.