Dual Action of Eeyarestatin 24 on Sec-Dependent Protein Secretion and Bacterial DNA
Eeyarestatin 24 (ES24) is really a promising new antibiotic with broad-spectrum activity. It shares structural similarity with nitrofurantoin (NFT), yet seems to possess a distinct and novel mechanism: ES24 was discovered to hinder SecYEG-mediated protein transport and membrane insertion in Gram-negative bacteria. However, possible additional targets have yet to be explored. Furthermore, its activity was particularly better against Gram-positive bacteria, that its mechanism of action had not investigated. We’ve used transcriptomic stress response profiling, phenotypic assays, and protein secretion Eeyarestatin 1 analyses to research the mode of action of ES24 in comparison to NFT while using Gram-positive model bacteria Bacillus subtilis and also have compared our findings to Gram-negative Escherichia coli. Here, we show the inhibition of Sec-dependent protein secretion in B. subtilis and furthermore provide evidence for DNA damage, most likely brought on by the generation of reactive derivatives of ES24. Interestingly, ES24 caused a gentle dissipation from the membrane potential, which brought to delocalization of cytokinetic proteins and subsequent cell elongation in E. coli. However, none of individuals effects were noticed in B. subtilis, therefore suggesting that ES24 displays distinct mechanistic variations regarding Gram-positive and Gram-negative bacteria. Despite its structural resemblance of NFT, ES24 profoundly differed within our phenotypic analysis, which means that it doesn’t share the NFT mechanism of generalized macromolecule and structural damage. Importantly, ES24 outperformed NFT in vivo inside a zebrafish embryo pneumococcal infection model. Our results claim that ES24 not just inhibits the Sec translocon, but additionally targets microbial DNA and, in Gram-negative bacteria, the cell membrane.