Souder Kendall, Beatty Emma J, McGovern Siena C, Whaby Michael, Young Emily, Pancake Jacob, Weekley Daron, Rice Justin, Primerano Donald A, Denvir James, Horzempa Joseph, Schmitt Deanna M
Department of Biomedical Sciences, West Liberty University, West Liberty, WV 26074, USA.
Department of Biomedical Sciences, Joan C. Edwards School of Medicine, Marshall University, Huntington, WV 25755, USA.
Antibiotics (Basel). 2021 Aug 17;10(8):992. doi: 10.3390/antibiotics10080992.
The phenoxazine dye resazurin exhibits bactericidal activity against the Gram-negative pathogens and . One resazurin derivative, resorufin pentyl ether, significantly reduces vaginal colonization by in a mouse model of infection. The narrow spectrum of bacteria susceptible to resazurin and its derivatives suggests these compounds have a novel mode of action. To identify potential targets of resazurin and mechanisms of resistance, we isolated mutants of subsp. live vaccine strain (LVS) exhibiting reduced susceptibility to resazurin and performed whole genome sequencing. The genes (FTL_0959) and (FTL_1306) were mutated in half of the 46 resazurin-resistant (RZR) strains sequenced. Complementation of select RZR LVS isolates with wild-type or partially restored sensitivity to resazurin. To further characterize the role of and in resazurin susceptibility, a deletion mutant, Δ, and disruption mutant, FTL_0959d, were generated. Both mutants were less sensitive to killing by resazurin compared to wild-type LVS with phenotypes similar to the spontaneous resazurin-resistant mutants. This study identified a novel role for two genes and in susceptibility to resazurin.
吩恶嗪染料刃天青对革兰氏阴性病原体和具有杀菌活性。刃天青的一种衍生物，试卤灵戊醚，在小鼠感染模型中显著减少了的阴道定植。对刃天青及其衍生物敏感的细菌谱较窄，表明这些化合物具有新的作用模式。为了确定刃天青的潜在靶点和耐药机制，我们分离了对刃天青敏感性降低的亚种活疫苗株（LVS）的突变体，并进行了全基因组测序。在测序的46株刃天青耐药（RZR）菌株中，有一半的菌株中基因（FTL_0959）和（FTL_1306）发生了突变。用野生型或对选定的RZR LVS分离株进行互补，部分恢复了对刃天青的敏感性。为了进一步表征和在刃天青敏感性中的作用，构建了一个缺失突变体Δ和一个破坏突变体FTL_0959d。与野生型LVS相比，这两个突变体对刃天青杀伤的敏感性均较低，其表型与自发的刃天青耐药突变体相似。本研究确定了两个基因和在对刃天青敏感性中的新作用。
