Corre Jacqueline, Louarn Jean-Michel
Laboratoire de Microbiologie et de Génétique Moléculaires, CNRS, 31062 Toulouse Cedex, France.
J Bacteriol. 2002 Jul;184(14):3801-7. doi: 10.1128/JB.184.14.3801-3807.2002.
Chromosome dimers in Escherichia coli are resolved at the dif locus by two recombinases, XerC and XerD, and the septum-anchored FtsK protein. Chromosome dimer resolution (CDR) is subject to strong spatiotemporal control: it takes place at the time of cell division, and it requires the dif resolution site to be located at the junction between the two polarized chromosome arms or replichores. Failure of CDR results in trapping of DNA by the septum and RecABCD recombination (terminal recombination). We had proposed that dif sites of a dimer are first moved to the septum by mechanisms based on local polarity and that normally CDR then occurs as the septum closes. To determine whether FtsK plays a role in the mobilization process, as well as in the recombination reaction, we characterized terminal recombination in an ftsK mutant. The frequency of recombination at various points in the terminus region of the chromosome was measured and compared with the recombination frequency on a xerC mutant chromosome with respect to intensity, the region affected, and response to polarity distortion. The use of a prophage excision assay, which allows variation of the site of recombination and interference with local polarity, allowed us to find that cooperating FtsK-dependent and -independent processes localize dif at the septum and that DNA mobilization by FtsK is oriented by the polarity probably due to skewed sequence motifs of the mobilized material.
大肠杆菌中的染色体二聚体通过两种重组酶XerC和XerD以及隔膜锚定的FtsK蛋白在dif位点处得以拆分。染色体二聚体拆分(CDR)受到严格的时空控制:它发生在细胞分裂时,并且需要dif拆分位点位于两个极化染色体臂或复制子之间的连接处。CDR失败会导致DNA被隔膜捕获并引发RecABCD重组(末端重组)。我们曾提出,二聚体的dif位点首先通过基于局部极性的机制移动到隔膜处,并且通常在隔膜闭合时发生CDR。为了确定FtsK是否在移动过程以及重组反应中发挥作用,我们对ftsK突变体中的末端重组进行了表征。测量了染色体末端区域各个点的重组频率,并将其与xerC突变体染色体上的重组频率在强度、受影响区域以及对极性扭曲的反应方面进行了比较。使用噬菌体切除试验,该试验允许改变重组位点并干扰局部极性,使我们发现协同的FtsK依赖性和非依赖性过程将dif定位在隔膜处,并且FtsK介导的DNA移动可能由于被移动物质的序列基序偏斜而由极性定向。