Microcolony formation by the opportunistic pathogen Pseudomonas aeruginosa requires pyruvate and pyruvate fermentation.
Microcolony formation by the opportunistic pathogen Pseudomonas aeruginosa requires pyruvate and pyruvate fermentation.
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DOI：
10.1111/mmi.12018
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发表时间：
2012-11
影响因子：
3.6
通讯作者：
Sauer K
中科院分区：
文献类型：
--
作者：
Petrova OE;Schurr JR;Schurr MJ;Sauer K
生物膜结构的一个标志是微菌落的存在。然而，人们对微集落形成的潜在机制知之甚少。在病原菌铜绿假单胞菌中，微集落的形成依赖于双组分调节剂MifR，mifR突变的生物膜表现出总体较薄的结构，缺乏微集落，而mifR的过表达导致超微集落形成。利用整体转录学和蛋白质组学的方法，我们证明了微克隆的形成与应激、氧气限制但电子丰富的条件有关，这表明应激反应机制的激活以及厌氧和发酵过程，特别是丙酮酸发酵。包括uspK、ACNA和LDHA在内的丙酮酸利用相关基因的失活以类似于mifR失活的方式减少了微集落的形成。此外，培养基中丙酮酸的耗尽会损害生物膜和微集落的形成，而添加丙酮酸则显著增加微集落的形成。在乳酸脱氢酶(LDHA)突变生物膜中加入丙酮酸或表达mifR不能恢复微集落形成，而丙酮酸的加入部分恢复了mifR突变生物膜中微集落的形成。相反，LDHA在mifR：：MAR中的表达完全恢复了该突变菌株的微集落形成。我们的发现表明，丙酮酸的发酵利用是对铜绿假单胞菌生物膜环境的微菌落特异性适应。
A hallmark of the biofilm architecture is the presence of microcolonies. However, little is known about the underlying mechanisms governing microcolony formation. In the pathogen Pseudomonas aeruginosa, microcolony formation is dependent on the two-component regulator MifR, with mifR mutant biofilms exhibiting an overall thin structure lacking microcolonies, and overexpression of mifR resulting in hyper-microcolony formation. Using global transcriptomic and proteomic approaches, we demonstrate that microcolony formation is associated with stressful, oxygen-limiting but electron-rich conditions, as indicated by the activation of stress response mechanisms and anaerobic and fermentative processes, in particular pyruvate fermentation. Inactivation of genes involved in pyruvate utilization including uspK, acnA and ldhA abrogated microcolony formation in a manner similar to mifR inactivation. Moreover, depletion of pyruvate from the growth medium impaired biofilm and microcolony formation, while addition of pyruvate significantly increased microcolony formation. Addition of pyruvate to or expression of mifR in lactate dehydrogenase (ldhA) mutant biofilms did not restore microcolony formation while addition of pyruvate partly restored microcolony formation in mifR mutant biofilms. In contrast, expression of ldhA in mifR::Mar fully restored microcolony formation by this mutant strain. Our findings indicate the fermentative utilization of pyruvate to be a microcolony-specific adaptation of the P. aeruginosa biofilm environment.
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影响因子：
3.7
作者：
Conibear TC;Collins SL;Webb JS
通讯作者：
Webb JS
影响因子：
3.2
作者：
Eschbach, M;Schreiber, K;Schobert, M
通讯作者：
Schobert, M
影响因子：
3.6
作者：
Dietrich, Lars E. P.;Price-Whelan, Alexa;Newman, Dianne K.
通讯作者：
Newman, Dianne K.
影响因子：
3.6
作者：
Allesen-Holm, M;Barken, KB;Tolker-Nielsen, T
通讯作者：
Tolker-Nielsen, T
影响因子：
3.2
作者：
CARLSON, CA;FERGUSON, LP;INGRAHAM, JL
通讯作者：
INGRAHAM, JL