MreB Drives De Novo Rod Morphogenesis in Caulobacter crescentus via Remodeling of the Cell Wall
MreB Drives De Novo Rod Morphogenesis in Caulobacter crescentus via Remodeling of the Cell Wall
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DOI：
10.1128/jb.01311-09
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发表时间：
2010-03-15
影响因子：
3.2
通讯作者：
Jacobs-Wagner, Christine
中科院分区：
文献类型：
--
作者：
Takacs, Constantin N.;Poggio, Sebastian;Jacobs-Wagner, Christine
MreB是细菌肌动蛋白样细胞骨架，是许多细菌物种杆状形态所必需的。MreB功能的破坏导致视杆形态和细胞变圆的损失。在这里，我们表明，广泛使用的MreB抑制剂A22导致MreB非依赖性生长抑制，其随药物浓度，培养基条件和测试的细菌种类而变化。MP265是一种A22结构类似物，对生长的毒性低于A22，但对破坏MreB细胞骨架同样有效。A22和MP265的作用通过培养基的碱性pH增强。利用这一知识和药物作用的快速可逆性，我们研究了在无毒条件下用MP265或A22预处理的柠檬形新月柄杆菌细胞中杆状的恢复。我们发现，药物去除后MreB功能的可逆恢复引起广泛的形态学变化，包括显著的细胞变薄伴随着伸长、细胞分支和外膜囊泡脱落。我们还对C.通过高效液相色谱法和质谱法分析了Crescentus肽聚糖的结构，并表明MreB功能恢复后MreB破坏和杆状恢复伴随着组成的显著变化。我们的研究结果提供了深入了解MreB在肽聚糖重塑和棒状形态发生中的功能，并表明MreB促进青霉素结合蛋白的转糖基酶活性。
MreB, the bacterial actin-like cytoskeleton, is required for the rod morphology of many bacterial species. Disruption of MreB function results in loss of rod morphology and cell rounding. Here, we show that the widely used MreB inhibitor A22 causes MreB-independent growth inhibition that varies with the drug concentration, culture medium conditions, and bacterial species tested. MP265, an A22 structural analog, is less toxic than A22 for growth yet equally efficient for disrupting the MreB cytoskeleton. The action of A22 and MP265 is enhanced by basic pH of the culture medium. Using this knowledge and the rapid reversibility of drug action, we examined the restoration of rod shape in lemon-shaped Caulobacter crescentus cells pretreated with MP265 or A22 under nontoxic conditions. We found that reversible restoration of MreB function after drug removal causes extensive morphological changes including a remarkable cell thinning accompanied with elongation, cell branching, and shedding of outer membrane vesicles. We also thoroughly characterized the composition of C. crescentus peptidoglycan by high-performance liquid chromatography and mass spectrometry and showed that MreB disruption and recovery of rod shape following restoration of MreB function are accompanied by considerable changes in composition. Our results provide insight into MreB function in peptidoglycan remodeling and rod shape morphogenesis and suggest that MreB promotes the transglycosylase activity of penicillin-binding proteins.