Chalmers R M, Kleckner N
Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
EMBO J. 1996 Sep 16;15(18):5112-22.
Transposon Tn10 and its component insertion sequence IS10 move by non-replicative transposition. We have studied the array of reaction intermediates and products in a high efficiency in vitro IS10/Tn10 transposition reaction. Synapsis of two transposon ends, followed by cleavage and strand transfer, can occur very efficiently irrespective of the relative locations and orientations of the two ends. The two participating ends can occur in inverted or direct orientation on the same molecule or, most importantly, on two different molecules. This behavior contrasts sharply with that of Mu, in which transposition is strongly biased in favor of inverted repeat synapsis. Mechanistically, the absence of discrimination amongst various end configurations implies that the architecture within the IS10/Tn10 synaptic complex is relatively simple, i.e. lacking any significant intertwining of component DNA strands. Biologically these observations are important because they suggest that the IS10 insertion sequence module has considerable flexibility in the types of DNA rearrangements that it can promote. Most importantly, it now seems highly probable that a single non-replicative IS10 element can promote DNA rearrangements usually attributed to replicative transposition, i.e. adjacent deletions and cointegrates, by utilizing transposon ends on two sister chromosomes. Other events which probably also contribute to the diversity of IS10/Tn10-promoted rearrangements are discussed.
转座子Tn10及其组成部分插入序列IS10通过非复制型转座进行移动。我们研究了高效体外IS10/Tn10转座反应中的一系列反应中间体和产物。两个转座子末端的联会,随后是切割和链转移,无论两个末端的相对位置和方向如何,都能非常高效地发生。参与的两个末端可以以反向或正向的方向出现在同一分子上,或者,最重要的是,出现在两个不同的分子上。这种行为与Mu的行为形成了鲜明对比,在Mu中,转座强烈偏向于反向重复联会。从机制上讲,对各种末端构型缺乏区分意味着IS10/Tn10突触复合体中的结构相对简单,即组成DNA链之间没有任何明显的缠绕。从生物学角度来看,这些观察结果很重要,因为它们表明IS10插入序列模块在其能够促进的DNA重排类型方面具有相当大的灵活性。最重要的是,现在看来极有可能单个非复制型IS10元件可以通过利用两条姐妹染色体上的转座子末端来促进通常归因于复制型转座的DNA重排,即相邻缺失和共合体。还讨论了可能也有助于IS10/Tn10促进的重排多样性的其他事件。