Co-translational protein aggregation and ribosome stalling as a novel broad-spectrum antibacterial mechanism

ABSTRACT: Drug- resistant bacteria pose are an urgent global threat health threatto global health, necessitatingrequiring the development of antibacterial compounds with a novel mode of action. Protein biosynthesis accounts for up to half of the energy expenditure of bacterial cells and consequently inhibiting the efficiency or fidelity of the bacterial ribosome is a major target of existing antibiotics. Here we describe an alternative mode of action to that affects the same process:; Allowing allowing translation to proceed but causing co-translational aggregation of the nascent chain. We found a peptide that acts via this mechanism and that is bactericidal against a wide range of pathogenic bacteria, including strains for which novel treatments are most urgently needed such as the ESKAPE pathogens, both in planktonic growth and in biofilms. Proteomics, transcriptomics and biochemical analyses of the inclusion bodies that form in bacteria upon peptide treatment reveal ribosomal proteins, as well the mRNAs and their cognate nascent chains of many proteins in amyloid-like structures assembled in polar inclusion bodies, with a bias for membrane proteins with a fold rich in long-range beta-sheet interactions. The bacterial ssrA ribosome rescue pathway is activated, and ribosome hibernation factors become enriched but fails to prevent this does not prevent the incorporation of the incorporation of hundreds of proteins into the inclusion bodies. Mechanistically, our results show that disrupting ribosomal quality controlprotein homeostasis via co-translational aggregation constitutes a potent target mechanism for the development ofdeveloping broad-spectrum antibacterials.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Escherichia Coli

SUBMITTER: Sebastien Carpentier  

LAB HEAD: Sebastien Carpentier

PROVIDER: PXD056779 | Pride | 2025-02-11

REPOSITORIES: pride