Novel dual regulators of Pseudomonas aeruginosa essential for productive biofilms and virulence.
Novel dual regulators of Pseudomonas aeruginosa essential for productive biofilms and virulence.
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DOI：
10.1111/mmi.14063
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发表时间：
2018-08
影响因子：
3.6
通讯作者：
Hoang TT
中科院分区：
文献类型：
--
作者：
Heacock-Kang Y;Zarzycki-Siek J;Sun Z;Poonsuk K;Bluhm AP;Cabanas D;Fogen D;McMillan IA;Chuanchuen R;Hoang TT
铜绿假单胞菌的基因调控网络是复杂的。由于基因组相对较大（6.2 Mb），有相当一部分基因被证明或预测为转录调节因子。许多这些调节剂已被证明在生物膜的形成和维持中发挥重要作用。在这项研究中，我们提出了一种新的转录调节因子，PA 1226，它调节铜绿假单胞菌的生物膜形成和毒力。编码该调节因子的基因突变消除了铜绿假单胞菌在体外产生生物膜的能力，而对铜绿假单胞菌的生长没有任何影响。这种调节剂也是在果蝇和BALB/c小鼠肺部感染模型中的体内适应性和发病机制所必需的。转录组分析表明，PA 1226调节许多重要的毒力基因/途径，包括那些参与藻酸盐，皮利，和LPS的生物合成。通过ChIP-seq鉴定由PA 1226直接调控的基因/操纵子和潜在的结合序列。通过电泳迁移率变动试验确认结合序列的尝试导致通过免疫共沉淀试验发现了共调节因子PA 1413。PA 1226和PA 1413显示协同结合其调节子的启动子区。提出了一个模型，总结了我们发现这种新的双调节系统。利用我们开创性的单个或几个原核细胞的总转录物扩增，我们以前已经发现了铜绿假单胞菌生物膜结构中数千个基因的空间依赖性表达。这一观察结果表明，一个复杂的细菌调控网络控制细菌的行为，这导致发现了一个新的双调节系统，PA 1226和PA 1413在这项工作中。在两种动物感染模型中，这种双调节系统在生物膜形成、体内适应性和发病机制中发挥重要作用。PA 1226和PA 1413合作调节几种基本毒力决定因素的基因表达，包括藻酸盐、LPS、绿脓杆菌素和皮利/菌毛生物合成操纵子。
Gene regulation network in Pseudomonas aeruginosa is complex. With a relatively large genome (6.2 Mb), there is a significant portion of genes that are proven or predicted to be transcriptional regulators. Many of these regulators have been shown to play important roles in biofilm formation and maintenance. In this study, we present a novel transcriptional regulator, PA1226, which modulates biofilm formation and virulence in P. aeruginosa. Mutation in the gene encoding this regulator abolished the ability of P. aeruginosa to produce biofilms in vitro, without any effect on the planktonic growth. This regulator is also essential for the in vivo fitness and pathogenesis in both Drosophila melanogaster and BALB/c mouse lung infection models. Transcriptome analysis revealed that PA1226 regulates many essential virulence genes/pathways, including those involved in alginate, pili, and LPS biosynthesis. Genes/operons directly regulated by PA1226 and potential binding sequences were identified via ChIP-seq. Attempts to confirm the binding sequences by electrophoretic mobility shift assay led to the discovery of a co-regulator, PA1413, via co-immunoprecipitation assay. PA1226 and PA1413 were shown to bind collaboratively to the promoter regions of their regulons. A model is proposed, summarizing our finding on this novel dual regulation system. Using our pioneering total transcript amplification of single or a few prokaryotic cells, we have previously uncovered the spatially dependent expressions of thousands of genes in the Pseudomonas aeruginosa biofilm architecture. This observation suggests a complex bacterial regulation network controlling bacterial behaviors, which led to the discovery of a novel dual-regulator system presented in this work, PA1226 and PA1413. This dual-regulator system is shown to play essential roles in biofilm formation, in vivo fitness and pathogenesis in two animal infection models. Collaboratively, PA1226 and PA1413 modulate the gene expressions of several essential virulence determinants, including alginate, LPS, pyoverdine, and pili/fimbriae biosynthesis operons.
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影响因子：
3.6
作者：
Kang Y;Lunin VV;Skarina T;Savchenko A;Schurr MJ;Hoang TT
通讯作者：
Hoang TT
影响因子：
3.2
作者：
Alm, RA;Bodero, AJ;Mattick, JS
通讯作者：
Mattick, JS
影响因子：
4
作者：
Ganne, Geraldine;Brillet, Karl;Schalk, Isabelle J.
通讯作者：
Schalk, Isabelle J.
影响因子：
3.6
作者：
Burrows, LL;Charter, DF;Lam, JS
通讯作者：
Lam, JS
影响因子：
4.4
作者：
De Kievit, TR;Gillis, R;Iglewski, BH
通讯作者：
Iglewski, BH