title: Auxotrophy-based High Throughput Screening assay for the identification of Bacillus subtilis stringent response inhibitors.
abstract: The stringent response is a central adaptation mechanism that allows bacteria to adjust their growth and metabolism according to environmental conditions. The functionality of the stringent response is crucial for bacterial virulence, survival during host invasion as well as antibiotic resistance and tolerance. Therefore, specific inhibitors of the stringent response hold great promise as molecular tools for disarming and pacifying bacterial pathogens. By taking advantage of the valine amino acid auxotrophy of the Bacillus subtilis stringent response-deficient strain, we have set up a High Throughput Screening assay for the identification of stringent response inhibitors. By screening 17,500 compounds, we have identified a novel class of antibacterials based on the 4-(6-(phenoxy)alkyl)-3,5-dimethyl-1H-pyrazole core. Detailed characterization of the hit compounds as well as two previously identified promising stringent response inhibitors - a ppGpp-mimic nucleotide Relacin and cationic peptide 1018 - showed that neither of the compounds is sufficiently specific, thus motivating future application of our screening assay to larger and more diverse molecular libraries.
authors: Department of Molecular Biology, Umeå University, Building 6K, 6L University Hospital Area, SE-901 87 Umeå, Sweden.|Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Building 6K and 6L, University Hospital Area, SE-901 87 Umeå, Sweden.
authors: Liis
authors: L
authors: Andresen
authors: Department of Molecular Biology, Umeå University, Building 6K, 6L University Hospital Area, SE-901 87 Umeå, Sweden.|Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Building 6K and 6L, University Hospital Area, SE-901 87 Umeå, Sweden.|University of Tartu, Institute of Technology, Nooruse 1, 50411 Tartu, Estonia.
authors: Vallo
authors: V
authors: Varik
authors: Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama, Saitama 338-8570, Japan.
authors: Yuzuru
authors: Y
authors: Tozawa
authors: Department of Molecular Biology, Umeå University, Building 6K, 6L University Hospital Area, SE-901 87 Umeå, Sweden.|Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Building 6K and 6L, University Hospital Area, SE-901 87 Umeå, Sweden.
authors: Steffi
authors: S
authors: Jimmy
authors: Laboratories for Chemical Biology Umeå (LCBU), Department of Chemistry, Umeå University, SE-901 87 Sweden.
authors: Stina
authors: S
authors: Lindberg
authors: University of Tartu, Institute of Technology, Nooruse 1, 50411 Tartu, Estonia.
authors: Tanel
authors: T
authors: Tenson
authors: Department of Molecular Biology, Umeå University, Building 6K, 6L University Hospital Area, SE-901 87 Umeå, Sweden.|Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Building 6K and 6L, University Hospital Area, SE-901 87 Umeå, Sweden.|University of Tartu, Institute of Technology, Nooruse 1, 50411 Tartu, Estonia.
authors: Vasili
authors: V
authors: Hauryliuk
mesh_terms: D000597:Amino Acids, Branched-Chain / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D001412:Bacillus subtilis / Q000187:drug effects* / Q000502:physiology; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000494:pharmacology; D004151:Dipeptides / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006159:Guanosine Tetraphosphate / Q000378:metabolism; D057166:High-Throughput Screening Assays; D011720:Pyrazoles / Q000737:chemistry / Q000494:pharmacology; D014633:Valine / Q000378:metabolism
chemical_list: D000597:Amino Acids, Branched-Chain; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004151:Dipeptides; D011720:Pyrazoles; C580565:relacin; D006159:Guanosine Tetraphosphate; D003849:Deoxyguanosine; D014633:Valine