Conjugative Interaction Induces Transposition of ISPst9 in Pseudomonas stutzeri AN10
Conjugative Interaction Induces Transposition of ISPst9 in Pseudomonas stutzeri AN10
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DOI：
10.1128/jb.01071-08
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发表时间：
2009-02-15
影响因子：
3.2
通讯作者：
Bosch, R.
中科院分区：
文献类型：
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作者：
Christie-Oleza, J. A.;Lanfranconi, M. P.;Bosch, R.
ISPst9是最近在降解萘的生物斯氏假单胞菌AN10中描述的一个类似ISL3的插入序列(IS)。在这篇文章中，我们描述了一种新的强IS调节刺激：ISPst9在所有的斯图泽里杆菌AN10细胞中与大肠杆菌接合作用后都被诱导了转座。因此，我们观察到，在所有通过接合获得遗传物质的Stutzeri AN10细胞中，ISPst9基因组的剂量和/或分布发生了变化。此外，当斯图尔氏杆菌AN10细胞与无质粒接合菌株E.coliS17-1 lambda(PIR)接触时，也观察到ISPst9转座，而当它们与E.coliDH5α(一种非接合菌株)接触时，则没有观察到ISPst9转座。分析了ISPst9转座的机制，证明了转座是通过从供体DNA中以保守的机制切除进行的，从而产生了侧翼DNA的3-10个碱基对的缺失。我们的结果表明，ISPst9发生了转位，形成了由IS和一个5-bp的插入DNA序列组成的双链DNA环状中间体，该序列可能来自ISPst9侧翼区域。并对IS环的形成动力学进行了描述。
ISPst9 is an ISL3-like insertion sequence (IS) that was recently described in the naphthalene-degrading organism Pseudomonas stutzeri strain AN10. In this paper we describe a novel strong IS regulation stimulus; transposition of ISPst9 is induced in all P. stutzeri AN10 cells after conjugative interaction with Escherichia coli. Thus, we observed that in all P. stutzeri AN10 cells that received genetic material by conjugation the ISPst9 genomic dose and/or distribution was changed. Furthermore, ISPst9 transposition was also observed when P. stutzeri AN10 cells were put in contact with the plasmidless conjugative strain E. coli S17-1 lambda(pir), but not when they were put in contact with E. coli DH5 alpha (a nonconjugative strain). The mechanism of ISPst9 transposition was analyzed, and transposition was shown to proceed by excision from the donor DNA using a conservative mechanism, which generated 3- to 10-bp deletions of the flanking DNA. Our results indicate that ISPst9 transposes, forming double-stranded DNA circular intermediates consisting of the IS and a 5-bp intervening DNA sequence probably derived from the ISPst9 flanking regions. The kinetics of IS circle formation are also described.