Hill T M, Kopp B J, Kuempel P L
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder 80309.
J Bacteriol. 1988 Feb;170(2):662-8. doi: 10.1128/jb.170.2.662-668.1988.
The terminus region of the Escherichia coli chromosome contains two sites that inhibit the progression of DNA replication forks. These termination sites, designated T1 and T2, are separated by 7.5 min (350 kilobases [kb]) on the genetic map and are located at the extremities of the terminus region. They demonstrate polarity (they stop replication forks traveling in one direction but not the other) and inhibit replication forks that have passed through and are about to leave the terminus. We have used deletion mutations in the terminus region to map the locations of T1 and T2 more accurately and to initiate studies on the mechanism of replication fork inhibition. We have narrowed the boundaries of T1 and T2 to 20 and 4 kb, respectively. T1 maps between kb 80 and 100 on the physical map of the terminus region (J. P. Bouché, J. Mol. Biol. 154:1-20, 1982), and T2 maps between kb 438 and 442. In addition, we report here that deletion of the region containing the T2 termination site inactivated T1. Supplying the T2 region on a plasmid restored T1 function, demonstrating that inhibition of replication at T1 requires a trans-acting factor which maps in the vicinity of termination site T2. We have called this newly identified terminus function the termination utilization substance (tus).
大肠杆菌染色体的末端区域包含两个抑制DNA复制叉前进的位点。这些终止位点,分别命名为T1和T2,在遗传图谱上相隔7.5分钟(350千碱基[kb]),位于末端区域的两端。它们表现出极性(它们阻止复制叉朝一个方向移动,但不阻止另一个方向),并抑制已经穿过并即将离开末端的复制叉。我们利用末端区域的缺失突变更精确地定位了T1和T2的位置,并开始研究复制叉抑制机制。我们已将T1和T2的边界分别缩小到20 kb和4 kb。T1位于末端区域物理图谱上的80 kb至100 kb之间(J. P. 布歇,《分子生物学杂志》154:1 - 20,1982),T2位于438 kb至442 kb之间。此外,我们在此报告,缺失包含T2终止位点的区域会使T1失活。在质粒上提供T2区域可恢复T1功能,表明T1处的复制抑制需要一个位于终止位点T2附近的反式作用因子。我们将这个新发现的末端功能称为终止利用物质(tus)。