Gilson E, Perrin D, Hofnung M
Unité de Programmation Moléculaire et Toxicologie Génétique, CNRS UA271 INSERM U163, Institut Pasteur, Paris, France.
Nucleic Acids Res. 1990 Jul 11;18(13):3941-52. doi: 10.1093/nar/18.13.3941.
Starting from a crude E. coli extract, two activities which specifically protect highly repetitive bacterial DNA sequences (called PU for Palindromic Unit or REP for Repetitive Extragenic Palindromic sequence) against a digestion with Exonuclease III have been purified. We show that one of these activities is due to the DNA polymerase I (Pol I). This constitutes the first indication for a specific interaction between Pol I and a duplex DNA. This interaction requires the presence of PU. It was confirmed and analyzed by native gel electrophoresis and DNase I footprinting experiments. The other activity contained at least five polypeptides. Its binding to PU DNA sequences was confirmed by native gel electrophoresis. Implications for the possible origin and functions of PU are discussed.
从粗制的大肠杆菌提取物开始,已经纯化出两种活性物质,它们能特异性保护高度重复的细菌DNA序列(称为回文单元的PU或重复基因外回文序列的REP)不被核酸外切酶III消化。我们发现其中一种活性物质是由于DNA聚合酶I(Pol I)。这是Pol I与双链DNA之间存在特异性相互作用的首个迹象。这种相互作用需要PU的存在。通过非变性凝胶电泳和DNase I足迹实验对其进行了证实和分析。另一种活性物质至少包含五种多肽。通过非变性凝胶电泳证实了其与PU DNA序列的结合。文中还讨论了PU可能的起源和功能。