Horiuchi T, Fujimura Y, Nishitani H, Kobayashi T, Hidaka M
National Institute for Basic Biology, Kyushu University, Fukuoka, Japan.
J Bacteriol. 1994 Aug;176(15):4656-63. doi: 10.1128/jb.176.15.4656-4663.1994.
In Escherichia coli, eight kinds of chromosome-derived DNA fragments (named Hot DNA) were found to exhibit homologous recombinational hotspot activity, with the following properties. (i) The Hot activities of all Hot DNAs were enhanced extensively under RNase H-defective (rnh) conditions. (ii) Seven Hot DNAs were clustered at the DNA replication terminus region on the E. coli chromosome and had Chi activities (H. Nishitani, M. Hidaka, and T. Horiuchi, Mol. Gen. Genet. 240:307-314, 1993). Hot activities of HotA, -B, and -C, the locations of which were close to three DNA replication terminus sites, the TerB, -A, and -C sites, respectively, disappeared when terminus-binding (Tau or Tus) protein was defective, thereby suggesting that their Hot activities are termination event dependent. Other Hot groups showed termination-independent Hot activities. In addition, at least HotA activity proved to be dependent on a Chi sequence, because mutational destruction of the Chi sequence on the HotA DNA fragment resulted in disappearance of the HotA activity. The HotA activity which had disappeared was reactivated by insertion of a new, properly oriented Chi sequence at the position between the HotA DNA and the TerB site. On the basis of these observations and positional and orientational relationships between the Chi and the Ter sequences, we propose a model in which the DNA replication fork blocked at the Ter site provides an entrance for the RecBCD enzyme into duplex DNA.
在大肠杆菌中,发现了8种源自染色体的DNA片段(命名为Hot DNA)表现出同源重组热点活性,具有以下特性。(i)在核糖核酸酶H缺陷(rnh)条件下,所有Hot DNA的Hot活性都大幅增强。(ii)7种Hot DNA聚集在大肠杆菌染色体上的DNA复制终止区,并且具有Chi活性(H. Nishitani、M. Hidaka和T. Horiuchi,《分子与普通遗传学》240:307 - 314,1993)。HotA、-B和-C的Hot活性,其位置分别靠近三个DNA复制终止位点,即TerB、-A和-C位点,当终止结合(Tau或Tus)蛋白有缺陷时,其Hot活性消失,从而表明它们的Hot活性依赖于终止事件。其他Hot组表现出不依赖于终止的Hot活性。此外,至少HotA活性被证明依赖于Chi序列,因为HotA DNA片段上Chi序列的突变破坏导致HotA活性消失。通过在HotA DNA与TerB位点之间的位置插入一个新的、正确定向的Chi序列,消失的HotA活性得以重新激活。基于这些观察结果以及Chi序列与Ter序列之间的位置和方向关系,我们提出一个模型,其中在Ter位点受阻的DNA复制叉为RecBCD酶进入双链DNA提供了一个入口。