Characterization of environmental regulators of Bordetella pertussis

Bordetella pertussis suppresses transcription of its virulence genes in response to specific environmental conditions, a response called modulation. The organism responds to high concentrations of SO4 and CIO4 ions, nicotinic acid, and nicotinic acid analogs in vitro; however, the in vivo modulator has not been identified. We investigated which chemical structures of the nicotinic acid molecule are important for modulation by testing various analogs for their ability to modulate. The ring nitrogen of nicotinic acid was not required, since benzoic acid was a modulator. In contrast, the carboxyl group was required, since derivatives like ethylnicotinate, 3-pyridylcarbinol, 3-acetyl pyridine, and 6-chloronicotinamide with altered carboxyl groups were not modulators. The planar ring structure or resonance in the ring was required for modulation, since nipecotic acid failed to modulate. The most potent modulators were nicotinic acid derivatives with electron-withdrawing substituents in the meta or para position relative to the carboxyl group. Relative hydrophilicity of substituents did not appear to contribute to modulation. Although these modulators elicited a clear biological response, the mechanism of modulation remains unclear, because no binding of the modulator 35SO4 or [14C]4-chlorobenzoic acid to whole B. pertussis was detected. However, modulation appears to involve a charge-charge interaction, since the response was blocked by chlorine ions.