Processing of Nonconjugative Resistance Plasmids by Conjugation Nicking Enzyme of Staphylococci.
Processing of Nonconjugative Resistance Plasmids by Conjugation Nicking Enzyme of Staphylococci.
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用葡萄球菌的接合切口酶加工非接合抗性质粒。
DOI：
10.1128/jb.00832-15
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发表时间：
2016
影响因子：
3.2
通讯作者：
Redinbo,MatthewR
中科院分区：
文献类型：
--
作者：
Pollet,RebeccaM;Ingle,JamesD;Hymes,JeffP;Eakes,ThomasC;Eto,KarinaYui;Kwong,StephenM;Ramsay,JoshuaP;Firth,Neville;Redinbo,MatthewR
金黄色葡萄球菌的抗微生物药物耐药性对人类健康的威胁日益严重。这种抗性通常编码在移动质粒上，如pSK41；然而，这些质粒的转移机制尚不清楚。在这项研究中，我们首先研究了松弛酶NES形成的关键蛋白质- dna相互作用，该酶启动和终止多药耐药质粒pSK41的转移。NES蛋白上的两个环，发夹环1和2，与pSK41理论区形成的DNA发夹形成广泛的接触，在这里我们建立了这些接触对于正确的DNA切割和体外完整的665个残基NES蛋白的定位是必不可少的。其次，pSK156和pCA347是非共轭性金黄色葡萄球菌质粒，它们含有与pSK41理论区相似的序列，但在预测形成DNA发夹的序列上不同。我们发现psk41编码的NES能够在体外结合、切割和修复这些非共轭质粒的理论序列。尽管pSK41可以动员含有其同源物的宿主质粒，但它不能动员含有pSK156和pCA347变异同源物的质粒，这表明一种类似于先前在pWBG749系统中被证明具有特异性的辅助蛋白也可能参与含有pSK41样同源物的质粒的传播。这些数据表明，最近描述的pWBG749家族质粒的共轭弛豫转运机制也适用于pSK41家族质粒，进一步强调了该机制在葡萄球菌质粒水平转移中的潜在意义。了解细菌（如金黄色葡萄球菌）的抗菌素耐药性转移机制是潜在地减缓抗菌素耐药性感染传播的重要一步。这项工作建立了金黄色葡萄球菌多抗性质粒pSK41通过其松弛酶NES转移的蛋白质- dna相互作用。这种酶也处理在许多以前认为不可传递的质粒上发现的变异蛋白样序列，表明这些质粒与一种未表征的辅助蛋白结合，可能通过一种弛豫机制水平转移。这些发现对我们理解葡萄球菌耐药质粒进化具有重要意义。
Antimicrobial resistance in Staphylococcus aureus presents an increasing threat to human health. This resistance is often encoded on mobile plasmids, such as pSK41; however, the mechanism of transfer of these plasmids is not well understood. In this study, we first examine key protein-DNA interactions formed by the relaxase enzyme, NES, which initiates and terminates the transfer of the multidrug resistance plasmid pSK41. Two loops on the NES protein, hairpin loops 1 and 2, form extensive contacts with the DNA hairpin formed at theoriTregion of pSK41, and here we establish that these contacts are essential for proper DNA cleavage and religation by the full 665-residue NES proteinin vitro. Second, pSK156 and pCA347 are nonconjugative Staphylococcus aureus plasmids that contain sequences similar to theoriTregion of pSK41 but differ in the sequence predicted to form a DNA hairpin. We show that pSK41-encoded NES is able to bind, cleave, and religate theoriTsequences of these nonconjugative plasmidsin vitro. Although pSK41 could mobilize a coresident plasmid harboring its cognateoriT, it was unable to mobilize plasmids containing the pSK156 and pCA347 variantoriTmimics, suggesting that an accessory protein like that previously shown to confer specificity in the pWBG749 system may also be involved in transmission of plasmids containing a pSK41-likeoriT. These data indicate that the conjugative relaxase intransmechanism recently described for the pWBG749 family of plasmids also applies to the pSK41 family of plasmids, further heightening the potential significance of this mechanism in the horizontal transfer of staphylococcal plasmids.IMPORTANCEUnderstanding the mechanism of antimicrobial resistance transfer in bacteria such as Staphylococcus aureus is an important step toward potentially slowing the spread of antimicrobial-resistant infections. This work establishes protein-DNA interactions essential for the transfer of the Staphylococcus aureus multiresistance plasmid pSK41 by its relaxase, NES. This enzyme also processed variantoriT-like sequences found on numerous plasmids previously considered nontransmissible, suggesting that in conjunction with an uncharacterized accessory protein, these plasmids may be transferred horizontally via a relaxase intransmechanism. These findings have important implications for our understanding of staphylococcal resistance plasmid evolution.
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影响因子：
14.9
作者：
O'Brien FG;Yui Eto K;Murphy RJ;Fairhurst HM;Coombs GW;Grubb WB;Ramsay JP
通讯作者：
Ramsay JP
影响因子：
3.7
作者：
Shen A;Lupardus PJ;Morell M;Ponder EL;Sadaghiani AM;Garcia KC;Bogyo M
通讯作者：
Bogyo M
影响因子：
3.6
作者：
通讯作者：
--
影响因子：
5.2
作者：
O'Brien, F. G.;Ramsay, J. P.;Grubb, W. B.
通讯作者：
Grubb, W. B.
影响因子：
15.9
作者：
Schommer NN;Gallo RL
通讯作者：
Gallo RL