Kidane Dawit, Graumann Peter L
Institut für Mikrobiologie, Fachbereich Biologie II, Universität Freiburg, Verfügungsgebäude, Stefan-Meier-Str. 19, 79104 Freiburg, Germany.
Cell. 2005 Jul 15;122(1):73-84. doi: 10.1016/j.cell.2005.04.036.
We have found that two DNA repair/recombination proteins localize differentially to the cell poles in competent Bacillus subtilis cells. RecA protein colocalized with competence protein ComGA, and its polar localization largely depended on ComGA and ComK activity, while RecN oscillated between the poles in a minute time frame, independent of any competence factor. Oscillation of RecN arrested upon addition of external DNA, suggesting that an interaction with incoming single-stranded (ss) DNA favors the localization of RecN at the pole containing the competence machinery. In agreement with this model, purified RecN protein showed ATP-dependent binding to ssDNA. Addition of DNA resulted in the formation of RecA threads emanating from the competence machinery. Our data show that in competent bacteria there exists a specifically positioned and dynamic ssDNA binding apparatus that accepts ssDNA taken up through the polar competence machinery and processes ssDNA for recombination with chromosomal DNA via extended RecA filaments.
我们发现,在感受态枯草芽孢杆菌细胞中,两种DNA修复/重组蛋白在细胞两极的定位存在差异。RecA蛋白与感受态蛋白ComGA共定位,其在两极的定位很大程度上依赖于ComGA和ComK的活性,而RecN在极短的时间内于两极之间振荡,与任何感受态因子无关。加入外源DNA后,RecN的振荡停止,这表明与进入的单链(ss)DNA相互作用有利于RecN定位于含有感受态机制的极。与该模型一致,纯化的RecN蛋白显示出对ssDNA的ATP依赖性结合。加入DNA导致从感受态机制发出RecA丝状物。我们的数据表明,在感受态细菌中存在一个特定定位且动态的ssDNA结合装置,该装置接受通过极性感受态机制摄取的ssDNA,并通过延伸的RecA丝状物处理ssDNA以与染色体DNA重组。