Becker Eric C, Meyer Richard J
Section of Molecular Genetics and Microbiology, School of Biology and Institute for Cellular and Molecular Biology, University of Texas at Austin, 78712, USA.
J Biol Chem. 2002 Apr 26;277(17):14575-80. doi: 10.1074/jbc.M110759200. Epub 2002 Feb 11.
MobA is a DNA strand transferase encoded by the plasmid R1162 and required for plasmid DNA processing during conjugal transfer. The smallest active fragment was identified using phage display and partial enzymatic digestion of the purified protein. This fragment, consisting of approximately the first 184 amino acids, is able to bind and cleave its normal DNA substrate, the origin of transfer (oriT). Smaller fragments having one of these activities were not obtained. An active intermediate consisting of MobA linked to DNA was isolated and used to show that a single molecule of MobA is sufficient to carry out all of the DNA processing steps during transfer. These results, along with those obtained earlier, point to a single large, active site in MobA that makes several different contacts along the oriT DNA strand.
MobA是一种由质粒R1162编码的DNA链转移酶,在接合转移过程中对质粒DNA加工至关重要。通过噬菌体展示和对纯化蛋白的部分酶切鉴定出最小的活性片段。该片段约由前184个氨基酸组成,能够结合并切割其正常DNA底物——转移起始点(oriT)。未获得具有这些活性之一的更小片段。分离出一种由与DNA相连的MobA组成的活性中间体,用于表明单个MobA分子足以在转移过程中执行所有的DNA加工步骤。这些结果,连同早期获得的结果,表明MobA中存在一个单一的大活性位点,该位点沿着oriT DNA链形成几种不同的接触。