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Integrated omics-based mechanistical insights into a membrane-targeting antibacterial

ABSTRACT: Here, we investigated HSI#6, a small-molecule antibacterial previously identified as a SecA activator, using integrated omics and functional assays. HSI#6 exhibits broad-spectrum bacteriostatic activity and acts through a dual-phase mechanism: transient activation of SecA-dependent secretion followed by membrane perturbation and global stress reprogramming. Time-resolved transcriptomics and proteomics revealed early activation of envelope stress regulons (Cpx, Rcs, Pho), efflux systems, and oxidative stress pathways, followed by suppression of ribosome biogenesis and central metabolism. Comparative analysis and biomarker-based principle component analysis (PCA) positioned HSI#6 within the envelope stress mechanistic space, closely aligned with membrane-active antibiotics yet displaying a distinct signature. Adaptive laboratory evolution (ALE) combined with whole-genome sequencing (WGS) revealed compensatory mutations in topoisomerase 1A gene (topA) and transcriptional regulators, without adaptive resistance emerged even under prolonged selection pressure. These findings establish HSI#6 as a mechanistically unique antibacterial targeting membrane homeostasis with low resistance potential.

INSTRUMENT(S):

ORGANISM(S): Escherichia Coli Bacteria

SUBMITTER: Haitham Abdallah

LAB HEAD: Lily Karamanou

PROVIDER: PXD075099 | Pride | 2026-05-22

REPOSITORIES: Pride

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			Action	DRS
	PCF000045_BW25113.7z	Other
	PCF000045_DIA-NN_output_and_imputed_matrix.xlsx	Xlsx
	PCF000045_Experimental_design_protein_ID.xlsx	Xlsx
	PCF000100_BW25113_Active_Kinetics.7z	Other
	PCF000100_DIA-NN_output_and_imputed_matrix.xlsx	Xlsx

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Integrated omics-based mechanistical insights into a membrane-targeting antibacterial

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