Deciphering the Antibacterial Mechanisms of 5-Fluorouracil in <i>Escherichia coli</i> through Biochemical and Transcriptomic Analyses
The emergence of carbapenem-resistant Gram-negative pathogens presents a clinical challenge in infection treatment, prompting the repurposing of existing drugs as an essential strategy to address this crisis. Although the anticancer drug 5-fluorouracil (5-FU) has been recognized for its antibacteria...
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| Main Authors: | , , , , , , |
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| Format: | Book |
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MDPI AG,
2024-06-01T00:00:00Z.
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| Summary: | The emergence of carbapenem-resistant Gram-negative pathogens presents a clinical challenge in infection treatment, prompting the repurposing of existing drugs as an essential strategy to address this crisis. Although the anticancer drug 5-fluorouracil (5-FU) has been recognized for its antibacterial properties, its mechanisms are not fully understood. Here, we found that the minimal inhibitory concentration (MIC) of 5-FU against <i>Escherichia coli</i> was 32-64 µg/mL, including strains carrying <i>bla</i><sub>NDM-5</sub>, which confers resistance to carbapenems. We further elucidated the antibacterial mechanism of 5-FU against <i>E. coli</i> by using genetic and biochemical analyses. We revealed that the mutation of uracil phosphoribosyltransferase-encoding gene <i>upp</i> increased the MIC of 5-FU against <i>E. coli</i> by 32-fold, indicating the role of the <i>upp</i> gene in 5-FU resistance. Additionally, transcriptomic analysis of <i>E. coli</i> treated with 5-FU at 8 µg/mL and 32 µg/mL identified 602 and 1082 differentially expressed genes involved in carbon and nucleic acid metabolism, DNA replication, and repair pathways. The biochemical assays showed that 5-FU induced bacterial DNA damage, significantly increased intracellular ATP levels and the NAD<sup>+</sup>/NADH ratio, and promoted reactive oxygen species (ROS) production. These findings suggested that 5-FU may exert antibacterial effects on <i>E. coli</i> through multiple pathways, laying the groundwork for its further development as a therapeutic candidate against carbapenem-resistant bacterial infections. |
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| Item Description: | 10.3390/antibiotics13060528 2079-6382 |