Raleigh E A, Kleckner N
J Mol Biol. 1984 Mar 15;173(4):437-61. doi: 10.1016/0022-2836(84)90390-5.
We have investigated the occurrence of multiple transposon-promoted chromosomal rearrangements in Escherichia coli K12 strains containing transposon Tn10. We show that a single Tn10 element, with its two closely spaced insertion sequence (IS10) elements, frequently gives rise to complex rearrangements that can be accounted for as the sum of two "classical" IS10 events. Using a strain containing differentially marked Tn10 elements at widely separated locations, we have investigated the possibility that IS10-promoted rearrangements occur in cell-wide "bursts", as expected if cells could occasionally undergo brief periods when all IS10 transposition events were activated, interspersed with longer periods of relative quiescence. We find no evidence for strong (greater than 60-fold), periodic cell-wide activation under our experimental conditions. The sensitivity of this experiment has been evaluated using an expression for the accumulation of double mutations in populations with heterogeneous, fluctuating mutation rates (see Appendix). We discuss several mechanisms by which two closely linked IS10 elements could undergo coupled double events without cell-wide activation: local activation of small chromosomal regions, periodic bursts of synthesis of cis-acting transposase protein, and/or a propensity for elements that have actually engaged in one rearrangement event to initiate a second successive event immediately thereafter. We favor the last possibility.
我们研究了含有转座子Tn10的大肠杆菌K12菌株中多个转座子促进的染色体重排的发生情况。我们发现,单个Tn10元件及其两个紧密间隔的插入序列(IS10)元件经常会引发复杂的重排,这些重排可以解释为两个“经典”IS10事件的总和。使用在广泛分离的位置含有差异标记的Tn10元件的菌株,我们研究了IS10促进的重排是否以全细胞“爆发”的形式发生的可能性,就像如果细胞偶尔会经历所有IS10转座事件都被激活的短暂时期,其间穿插着较长的相对静止期时所预期的那样。在我们的实验条件下,我们没有发现强烈的(大于60倍)、周期性的全细胞激活的证据。使用具有异质、波动突变率的群体中双突变积累的表达式评估了该实验的敏感性(见附录)。我们讨论了几种机制,通过这些机制,两个紧密相连的IS10元件可以在没有全细胞激活的情况下发生偶联双事件:小染色体区域的局部激活、顺式作用转座酶蛋白合成的周期性爆发和/或实际上已经参与一次重排事件的元件紧接着立即引发第二次连续事件的倾向。我们倾向于最后一种可能性。