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首页> 外文期刊>Journal of Molecular Biology >DNA sequence bias during Tn5 transposition.
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DNA sequence bias during Tn5 transposition.

机译：Tn5转座过程中的DNA序列偏倚。

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摘要

Transposition is one of the primary mechanisms causing genome instability. This phenomenon is mechanistically related to other DNA rearrangements such as V(D)J recombination and retroviral DNA integration. In the Tn5 system, only one protein, the transposase (Tnp), is required for all of the catalytic steps involved in transposon movement. The complexity involved in moving multiple DNA strands within one active site suggests that, in addition to the specific contacts maintained between Tnp and its recognition sequence, Tnp also interacts with the flanking DNA sequence. Here, we demonstrate that Tnp interacts with the donor DNA region. Tnp protects the donor DNA from DNase I digestion, suggesting that Tnp is in contact with, or otherwise distorts, the donor DNA during synapsis. In addition, changes in the donor DNA sequence within this region alter the affinity of Tnp for DNA by eightfold during synapsis. In vitro selection for more stable synaptic complexes reveals an A/T sequence bias for this region.We further show that certain donor DNA sequences, which favor synapsis, also appear to serve as hot spots for strand transfer. The TTATA donor sequence represents the best site. Most surprising is the fact that this sequence is found within the Tnp recognition sequence. Preference for insertion into a site within the Tnp recognition sequence would effectively inactivate one copy of the element and form clusters of the Tn5 transposon. In addition, the fact that several donor DNA sequences, which favor synapsis, appear to serve as hot spots for transposon insertion suggest that similar criteria may exist for Tnp-donor DNA and Tnp-target DNA interactions.
机译：转座是引起基因组不稳定的主要机制之一。这种现象在机械上与其他DNA重排有关，例如V（D）J重组和逆转录病毒DNA整合。在Tn5系统中，与转座子运动有关的所有催化步骤仅需要一种蛋白质，即转座酶（Tnp）。在一个活性位点内移动多条DNA链所涉及的复杂性表明，除了在Tnp及其识别序列之间保持特异性接触外，Tnp还与侧翼DNA序列相互作用。在这里，我们证明Tnp与供体DNA区域相互作用。 Tnp保护供体DNA免受DNase I消化，表明Tnp在突触过程中与供体DNA接触或扭曲。另外，在突触过程中，该区域内供体DNA序列的改变使Tnp对DNA的亲和力改变了八倍。体外筛选更稳定的突触复合物揭示了该区域的A / T序列偏向性。我们进一步表明，某些有利于突触的供体DNA序列似乎也充当链转移的热点。 TTATA供体序列代表最佳位点。最令人惊讶的是，在Tnp识别序列中发现了该序列。优选插入Tnp识别序列内的位点将有效地灭活该元件的一个拷贝并形成Tn5转座子的簇。另外，一些有利于突触的供体DNA序列似乎充当转座子插入的热点这一事实表明，对于Tnp-供体DNA和Tnp-靶标DNA相互作用可能存在相似的标准。

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