Stietz Maria S, Lopez Christina, Osifo Osasumwen, Tolmasky Marcelo E, Cardona Silvia T
a Department of Microbiology, University of Manitoba, Winnipeg, MB R3T 2N2, Canada.
b Department of Biological Science, Center for Applied Biotechnology Studies, California State University Fullerton, Fullerton, CA 92831-3599, USA.
Can J Microbiol. 2017 Oct;63(10):857-863. doi: 10.1139/cjm-2017-0350. Epub 2017 Aug 17.
There are hundreds of essential genes in multidrug-resistant bacterial genomes, but only a few of their products are exploited as antibacterial targets. An example is the electron transfer flavoprotein (ETF), which is required for growth and viability in Burkholderia cenocepacia. Here, we evaluated ETF as an antibiotic target for Burkholderia cepacia complex (Bcc). Depletion of the bacterial ETF during infection of Caenorhabditis elegans significantly extended survival of the nematodes, proving that ETF is essential for survival of B. cenocepacia in this host model. In spite of the arrest in respiration in ETF mutants, the inhibition of etf expression did not increase the formation of persister cells, when treated with high doses of ciprofloxacin or meropenem. To test if etf translation could be inhibited by RNA interference, antisense oligonucleotides that target the etfBA operon were synthesized. One antisense oligonucleotide was effective in inhibiting etfB translation in vitro but not in vivo, highlighting the challenge of reduced membrane permeability for the design of drugs against B. cenocepacia. This work contributes to the validation of ETF of B. cenocepacia as a target for antibacterial therapy and demonstrates the utility of a C. elegans liquid killing assay to validate gene essentiality in an in vivo infection model.
多重耐药细菌基因组中有数百个必需基因,但只有少数基因产物被用作抗菌靶点。电子传递黄素蛋白(ETF)就是一个例子,它是洋葱伯克霍尔德菌生长和存活所必需的。在此,我们评估了ETF作为洋葱伯克霍尔德菌复合体(Bcc)的抗生素靶点。在秀丽隐杆线虫感染期间消耗细菌ETF可显著延长线虫的存活时间,证明ETF对该宿主模型中洋葱伯克霍尔德菌的存活至关重要。尽管ETF突变体的呼吸受阻,但在用高剂量环丙沙星或美罗培南治疗时,抑制etf表达并未增加持留菌细胞的形成。为了测试RNA干扰是否能抑制etf翻译,合成了靶向etfBA操纵子的反义寡核苷酸。一种反义寡核苷酸在体外有效抑制etfB翻译,但在体内无效,这凸显了针对洋葱伯克霍尔德菌设计药物时膜通透性降低的挑战。这项工作有助于验证洋葱伯克霍尔德菌的ETF作为抗菌治疗靶点的有效性,并证明了秀丽隐杆线虫液体杀伤试验在体内感染模型中验证基因必需性的实用性。