Li Yongfang, Youngren Brenda, Sergueev Kirill, Austin Stuart
Gene Regulation and Chromosome Biology Laboratory, National Cancer Institute, CCR, NCI-Frederick, Frederick, Maryland 21702-1201, USA.
Mol Microbiol. 2003 Nov;50(3):825-34. doi: 10.1046/j.1365-2958.2003.03746.x.
We studied the segregation of the replication terminus of the Escherichia coli chromosome by time-lapse and still photomicroscopy. The replicated termini lie together at the cell centre. They rapidly segregate away from each other immediately before cell division. At fast growth rate, the copies move progressively and quickly toward the centres of the new-born cells. At slow growth rate, the termini usually remain near the inner cell pole and migrate to the cell centre in the middle of the cell cycle. A terminus domain of about 160kb, roughly centred on the dif recombination site, segregated as a unit at cell division. Sequences outside this domain segregated before division, giving two separate foci in predivision cells. Resolution of chromosome dimers via the terminus dif site requires the XerC recombinase and an activity of the FtsK protein that is thought to align the dif sequences at the cell centre. We found that anchoring of the termini at the cell centre and proper segregation at cell division occurred normally in the absence of recombination via the XerC recombinase. Anchoring and proper segregation were, however, frequently disrupted when the C-terminal domain of FtsK was truncated.
我们通过延时和静态显微镜观察研究了大肠杆菌染色体复制终点的分离情况。复制后的终点位于细胞中心。它们在细胞分裂前迅速彼此分离。在快速生长速率下,复制产物逐渐且迅速地移向新生细胞的中心。在缓慢生长速率下,终点通常保持在细胞内极附近,并在细胞周期中期迁移至细胞中心。一个约160kb的终点区域,大致以dif重组位点为中心,在细胞分裂时作为一个整体分离。该区域外的序列在分裂前分离,在分裂前细胞中形成两个独立的焦点。通过终点dif位点解决染色体二聚体问题需要XerC重组酶和FtsK蛋白的一种活性,该活性被认为可使dif序列在细胞中心对齐。我们发现,在缺乏通过XerC重组酶进行重组的情况下,终点在细胞中心的锚定以及在细胞分裂时的正确分离正常发生。然而,当FtsK的C末端结构域被截短时,锚定和正确分离经常受到干扰。