The Salmonella LysR Family Regulator RipR Activates the SPI-13-Encoded Itaconate Degradation Cluster

• Published in: Infection and immunity Vol. 88; no. 10
• Main Authors: Hersch, Steven J, Navarre, William Wiley
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 18.09.2020
• Subjects: Animals; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Cell Line; Gene Expression Regulation, Bacterial - drug effects; Genomic Islands - genetics; Humans; Hydrogen-Ion Concentration; Macrophages - metabolism; Macrophages - microbiology; Mice; Molecular Pathogenesis; Operon; Phagosomes - metabolism; Phagosomes - microbiology; Promoter Regions, Genetic; Salmonella typhimurium - drug effects; Salmonella typhimurium - genetics; Salmonella typhimurium - metabolism; Succinates - metabolism; Succinates - pharmacology; Transcription Factors - genetics; Transcription Factors - metabolism; Salmonella; host-pathogen interactions; macrophages; itaconate; acid resistance; gene regulation; pathogenicity islands; biosensors; metabolism; promoters
• ISSN: 0019-9567; 1098-5522
• DOI: 10.1128/IAI.00303-20

Itaconate is a dicarboxylic acid that inhibits the isocitrate lyase enzyme of the bacterial glyoxylate shunt. Activated macrophages have been shown to produce itaconate, suggesting that these immune cells may employ this metabolite as a weapon against invading bacteria. Here, we demonstrate that , itaconate can exhibit bactericidal effects under acidic conditions similar to the pH of a macrophage phagosome. In parallel, successful pathogens, including , have acquired a genetic operon encoding itaconate degradation proteins, which are induced heavily in macrophages. We characterized the regulation of this operon by the neighboring gene in specific response to itaconate. Moreover, we developed an itaconate biosensor based on the operon promoter that can detect itaconate in a semiquantitative manner and, when combined with the gene, is sufficient for itaconate-regulated expression in Using this biosensor with fluorescence microscopy, we observed bacteria responding to itaconate in the phagosomes of macrophages and provide additional evidence that gamma interferon stimulates macrophage itaconate synthesis and that J774 mouse macrophages produce substantially more itaconate than the human THP-1 monocyte cell line. In summary, we examined the role of itaconate as an antibacterial metabolite in mouse and human macrophages, characterized the regulation of 's defense against it, and developed it as a convenient itaconate biosensor and inducible promoter system.