Dual Action of Eeyarestatin 24 on Sec-Dependent Protein Secretion and Bacterial DNA
Artikel i vetenskaplig tidskrift, 2023

Eeyarestatin 24 (ES24) is a promising new antibiotic with broad-spectrum activity. It shares structural similarity with nitrofurantoin (NFT), yet appears to have a distinct and novel mechanism: ES24 was found to inhibit SecYEG-mediated protein transport and membrane insertion in Gram-negative bacteria. However, possible additional targets have not yet been explored. Moreover, its activity was notably better against Gram-positive bacteria, for which its mechanism of action had not yet been investigated. We have used transcriptomic stress response profiling, phenotypic assays, and protein secretion analyses to investigate the mode of action of ES24 in comparison with NFT using the Gram-positive model bacterium Bacillus subtilis and have compared our findings to Gram-negative Escherichia coli. Here, we show the inhibition of Sec-dependent protein secretion in B. subtilis and additionally provide evidence for DNA damage, probably caused by the generation of reactive derivatives of ES24. Interestingly, ES24 caused a gradual dissipation of the membrane potential, which led to delocalization of cytokinetic proteins and subsequent cell elongation in E. coli. However, none of those effects were observed in B. subtilis, thereby suggesting that ES24 displays distinct mechanistic differences with respect to Gram-positive and Gram-negative bacteria. Despite its structural similarity to NFT, ES24 profoundly differed in our phenotypic analysis, which implies that it does not share the NFT mechanism of generalized macromolecule and structural damage. Importantly, ES24 outperformed NFT in vivo in a zebrafish embryo pneumococcal infection model. Our results suggest that ES24 not only inhibits the Sec translocon, but also targets bacterial DNA and, in Gram-negative bacteria, the cell membrane.

in vivo efficacy


protein secretion


mechanism of action

eeyarestatin 24


Ann-Britt Schäfer

Chalmers, Life sciences, Kemisk biologi

Maurice Steenhuis

Vrije Universiteit Amsterdam

Kin Ki Jim

Universiteit Van Amsterdam

Amsterdam institute for Infection and Immunity

Jolanda Neef

Rijksuniversiteit Groningen

Sarah O’Keefe

University of Manchester

Roger C. Whitehead

University of Manchester

Eileithyia Swanton

University of Manchester

Biwen Wang

Swammerdam Institute for Life Sciences

Sven Halbedel

Robert Koch Institut

Otto von Guericke Universitaet Magdeburg

Stephen High

University of Manchester

Jan Maarten van Dijl

Rijksuniversiteit Groningen

Joen Luirink

Vrije Universiteit Amsterdam

Michaela Wenzel

Chalmers, Life sciences, Kemisk biologi

ACS Infectious Diseases

2373-8227 (eISSN)

Vol. 9 2 253-269


Biokemi och molekylärbiologi






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