iTP-Seq reveals how the broad-spectrum protein synthesis inhibitor sparsomycin remodels the bacterial translation landscape in vitro
Sparsomycin is a broad-spectrum inhibitor of protein synthesis with activity across all three domains of life. In this study, we used inverse toeprinting coupled to next-generation sequencing (iTP-Seq) to determine how sparsomycin remodels the bacterial translation landscape in vitro. Our data reveals that 0.1 mM sparsomycin blocks ribosomes engaged in the synthesis of a majority of nascent peptide sequences, but nevertheless displays some degree of context-specificity in its ability to inhibit translation. By shedding new light on the mode of action of sparsomycin on the bacterial ribosome, our work provides a framework for the design of sparsomycin derivatives with improved potency and bacterial specificity.