Inhibition of autotransporter biogenesis by small molecules. Issue 1 (3rd May 2019)

• Record Type: Journal Article
• Authors: Steenhuis, Maurice; Abdallah, Abdallah M.; de Munnik, Sabrina M.; Kuhne, Sebastiaan; Sterk, Geert‐Jan; van den Berg van Saparoea, Bart; Westerhausen, Sibel; Wagner, Samuel; van der Wel, Nicole N.; Wijtmans, Maikel; van Ulsen, Peter; Jong, Wouter S. P.; Luirink, Joen
• Is Part Of: Molecular microbiology. Volume 112:Issue 1(2019)
• Page Start: 81
• Page End: 98
• Publication Date: 2019-05-03
• Subjects: Molecular microbiology -- Periodicals; 572.829
• DOI: 10.1111/mmi.14255
• Languages: English
• ISSNs: 0950-382X

Abstract

Summary: Disarming pathogens by targeting virulence factors is a promising alternative to classic antibiotics. Many virulence factors in Gram‐negative bacteria are secreted via the autotransporter (AT) pathway, also known as Type 5 secretion. These factors are secreted with the assistance of two membrane‐based protein complexes: Sec and Bam. To identify inhibitors of the AT pathway, we used transcriptomics analysis to develop a fluorescence‐based high‐throughput assay that reports on the stress induced by the model AT hemoglobin protease (Hbp) when its secretion across the outer membrane is inhibited. Screening a library of 1600 fragments yielded the compound VUF15259 that provokes cell envelope stress and secretion inhibition of the ATs Hbp and Antigen‐43. VUF15259 also impairs β‐barrel folding activity of various outer membrane proteins. Furthermore, we found that mutants that are compromised in outer membrane protein biogenesis are more susceptible to VUF15259. Finally, VUF15259 induces the release of vesicles that appear to assemble in short chains. Taken together, VUF15259 is the first reported compound that inhibits AT secretion and our data are mostly consistent with VUF15259 interfering with the Bam‐complex as potential mode of action. The validation of the presented assay incites its use to screen larger compound libraries with drug‐like compounds. Abstract : A promising new category of antibiotics are molecules that interfere with virulence factors. Many virulence factors are transported across the bacterial cell envelope into the host by the autotransporter (AT) pathway. We developed a high‐throughput assay that reports on compromised AT secretion. Screening a compound library resulted in an inhibitor of AT secretion that also interfered with biogenesis of β‐barrel outer membrane proteins.