title: Isolation and characterization of inhibitors of the essential histidine kinase, YycG in Bacillus subtilis and Staphylococcus aureus.
abstract: The set of sensor kinase YycG and response regulator YycF is the only essential two-component system (TCS) in Bacillus subtilis and Staphylococcus aureus. We have developed a screening method for antibacterial agents that inhibit YycG, the essential histidine kinase (HK). To increase screening sensitivity, a temperature-sensitive yycF mutant (CNM2000) of B. subtilis with super-sensitivity to HK inhibitors was constructed, which was used for the screening of acetone extracts from 4000 microbes. A total of 11 samples showed higher sensitivity to CNM2000 than to wild-type parent 168, and seven of those were characterized to be potent inhibitors against autophosphorylation of YycG. One sample compound was purified and identified as aranorosinol B, a known antibacterial agent against Gram-positive bacteria including B. subtilis and S. aureus. Aranorosinol B inhibited YycG from both B. subtilis and S. aureus with a half-maximum inhibitory concentration (IC50) of 223 and 211 microM, respectively.
authors.affiliation: Department of Bioscience and Biotechnology, Graduate School of Agriculture, Kinki University, 3327-204, Nakamachi, Nara 631-8505, Japan.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Watanabe
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Hashimoto
authors.forename: Kaneyoshi
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Kiyo
authors.initials: K
authors.lastname: Hirao
authors.forename: Akira
authors.initials: A
authors.lastname: Ishihama
authors.forename: Motohiro
authors.initials: M
authors.lastname: Hino
authors.forename: Ryutaro
authors.initials: R
authors.lastname: Utsumi
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001412:Bacillus subtilis / Q000737:chemistry*; D001483:Base Sequence; D017931:DNA Primers; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D000071677:Histidine Kinase; D010766:Phosphorylation; D010957:Plasmids; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D013211:Staphylococcus aureus / Q000737:chemistry*
chemical_list: D000900:Anti-Bacterial Agents; D017931:DNA Primers; D004791:Enzyme Inhibitors; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D000071677:Histidine Kinase