Cameron R K, Ulycznyj P I, DuBow M S
Department of Microbiology and Immunology, McGill University, Montreal, Quebec, Canada.
Res Microbiol. 1995 Oct;146(8):601-16. doi: 10.1016/0923-2508(96)81059-x.
The transposable bacteriophage Mu and the mobile genetic elements Tn3 and IS101 replicatively transpose to random target sites, produce 5 bp target site duplications, and contain the sequence 5'-PuCGAAAPu-3' starting at bp 21 from their ends. The presence of these shared characteristics, plus the fact that Mu transposase can specifically bind to the termini of Tn3 and IS101 in vitro, suggests that the elements may be evolutionarily conserved and retain some functional capacity to transpose each other's DNA. To examine this proposition, in vivo transposition-mating assays were performed and demonstrated that Mu transposase stimulated the formation of recA-independent recombination products between Tn3kan- or IS101-containing plasmids and a target plasmid (pOX38cam) up to 200-fold. However, when transferred to recA+ hosts, these recA-independent products yielded resolution products suggestive of illegitimate recombination, as similar recombination and resolution products were generated, at reduced frequencies, in the absence of Mu transposase. Thus, Mu transposase may stimulate a host-mediated, recA-independent illegitimate recombination reaction. As adjacent pSC101 sequences, including a formerly unknown but functional IHF site (bp 2238-2251), were required for Mu transposase-stimulated IS101 illegitimate recombination, IHF may be one of the putative host factors involved in these recombination reactions.
可转座噬菌体Mu以及移动遗传元件Tn3和IS101以复制方式转座到随机靶位点,产生5 bp的靶位点重复序列,并在距其末端21 bp处含有5'-PuCGAAAPu-3'序列。这些共同特征的存在,加上Mu转座酶在体外可特异性结合Tn3和IS101末端这一事实,表明这些元件在进化上可能是保守的,并保留了一些转座彼此DNA的功能能力。为了验证这一观点,进行了体内转座交配试验,结果表明Mu转座酶可将含Tn3kan或IS101的质粒与靶质粒(pOX38cam)之间recA非依赖性重组产物的形成刺激高达200倍。然而,当转移到recA+宿主中时,这些recA非依赖性产物产生了提示非法重组的解离产物,因为在没有Mu转座酶的情况下,也会以较低频率产生类似的重组和解离产物。因此,Mu转座酶可能刺激宿主介导的、recA非依赖性的非法重组反应。由于Mu转座酶刺激的IS101非法重组需要相邻的pSC101序列,包括一个以前未知但有功能的整合宿主因子(IHF)位点(bp 2238-2251),因此IHF可能是参与这些重组反应的假定宿主因子之一。