Tn5/IS50 target recognition
Tn5/IS50 target recognition
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DOI：
10.1073/pnas.95.18.10716
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发表时间：
1998-09-01
影响因子：
11.1
通讯作者：
Reznikoff, WS
中科院分区：
文献类型：
--
作者：
Goryshin, IY;Miller, JA;Reznikoff, WS
该通信通过在两个相对较小的DNA区域（每个小于1 kb）中使用大量TN5和IS50插入来报告TN5/IS50目标位点选择的分析。对于两个区域，都根据体外和体内转置事件收集了数据。由于数据集是一致的，并且转座酶是体外唯一存在的蛋白质，因此这表明目标选择是仅转座酶的特性，因此似乎有两个有关目标选择的因素。通过分析所有插入位点，推断出一个靶标共识序列，该序列大概反映了单个TN5/IS50结合的转座酶量子的目标选择。共识TN5/IS50目标站点是A-GntyWranc-T。但是，我们观察到独立的插入位点倾向于形成位置紧密的插入（簇）组，并且插入经常以5 bp的周期性进行间隔。这表明TA5/IS50靶标的选择是通过与DNA结合的两个以上的转座酶素的促进的，因此，要使位点成为一个好的靶标，重叠的相邻DNA也应该是一个好的目标，合成目标序列也是设计和用于测试和确认此模型。
This communication reports an analysis of Tn5/IS50 target site selection by using an extensive collection of Tn5 and IS50 insertions in two relatively small regions of DNA (less than 1 kb each). For both regions data were collected resulting from in vitro and in vivo transposition events. Since the data sets are consistent and transposase was the only protein present in vitro, this demonstrates that target selection is a property of only transposase, There appear to be two factors governing target selection. A target consensus sequence, which presumably reflects the target selection of individual pairs of Tn5/IS50 bound transposase protomers, was deduced by analyzing all insertion sites. The consensus Tn5/IS50 target site is A-GNTYWRANC-T. However, we observed that independent insertion sites tend to form groups of closely located insertions (clusters), and insertions very often were spaced in a 5-bp periodic fashion. This suggests that Ta5/IS50 target selection is facilitated by more than two transposase protomers binding to the DNA, and, thus, for a site to be a good target, the overlapping neighboring DNA should be a good target, too, Synthetic target sequences were designed and used to test and confirm this model.