Reverse engineering antibiotic sensitivity in a multidrug-resistant Pseudomonas aeruginosa isolate.
Reverse engineering antibiotic sensitivity in a multidrug-resistant Pseudomonas aeruginosa isolate.
复制标题
多重耐药铜绿假单胞菌分离株的逆向工程抗生素敏感性。
DOI：
10.1128/aac.01640-05
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Gill,RyanT
中科院分区：
文献类型：
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作者：
Struble,JulieM;Gill,RyanT
抗生素耐药性是一个普遍和日益严重的临床问题。我们描述了一个评估的逆向工程方法，以确定细胞机制和基因，可以被操纵，以增加抗生素的敏感性，在resistantPseudomonasaerobososisolate。我们首先用化学方法使一种广泛耐药的分离株发生突变。aerodysaccharide和筛选对氨基糖苷类阿米卡星敏感性增加的突变体，然后对突变体和野生型菌株进行全基因组转录谱分析，以表征由于突变而发生的全局变化。然后，我们进行了一系列测定，以表征突变株敏感性增加的机制。我们报告了四个主要结果：（i）相对于增加耐药性的突变频率，增加敏感性的突变发生频率较高（10 - 2（10 - 5至10 - 10），并且发生频率比单个点突变的频率高104;（ii）在敏感突变体中转录谱被改变，导致总体表达模式与敏感实验室菌株PAO 1的表达模式比亲本抗性菌株的表达模式更相似;（iii）从转录谱中发现的基因在与细胞膜通透性和氨基糖苷修饰相关的表达编码功能中具有更显著的变化，这两者都是已知的氨基糖苷类耐药机制;最后，（iv）即使我们没有确定特定的突变位点，但几种不同的后续MIC测定表明，敏感突变体之间导致敏感性增加的突变不同。
Antibiotic resistance is a pervasive and growing clinical problem. We describe an evaluation of a reverse engineering approach for identifying cellular mechanisms and genes that could be manipulated to increase antibiotic sensitivity in a resistantPseudomonas aeruginosaisolate. We began by chemically mutating a broadly resistant isolate ofP. aeruginosaand screening for mutants with increased sensitivity to the aminoglycoside amikacin, followed by performing whole-genome transcriptional profiling of the mutant and wild-type strains to characterize the global changes occurring as a result of the mutations. We then performed a series of assays to characterize the mechanisms involved in the increased sensitivity of the mutant strains. We report four primary results: (i) mutations that increase sensitivity occur at a high frequency (10−2) relative to the frequency of those that increase resistance (10−5to 10−10) and occur at a frequency 104higher than the frequency of a single point mutation; (ii) transcriptional profiles were altered in sensitive mutants, resulting in overall expression patterns more similar to those of the sensitive laboratory strain PAO1 than those of the parental resistant strain; (iii) genes found from transcriptional profiling had the more dramatic changes in expression-encoded functions related to cellular membrane permeability and aminoglycoside modification, both of which are known aminoglycoside resistance mechanisms; and finally, (iv) even though we did not identify the specific sites of mutation, several different follow-up MIC assays suggested that the mutations responsible for increased sensitivity differed between sensitive mutants.