Transcriptional regulation of drug efflux genes by EvgAS, a two-component system in Escherichia coli

1 Department of Bioscience and Biotechnology, Graduate School of Agriculture of Kinki University, 3327-204, Nakamachi, Nara 631-8505, Japan 2 Research and Education Center for Genetic Information, Nara Institute of Science and Technology, Ikoma 630-0101, Japan 3 Department of Bioengineering, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta 4259, Midori-ku, Yokohama 226-0027, Japan 4 Division of Molecular Biology, Nippon Institute for Biological Science, Ome, Tokyo 190-0024, Japan Correspondence Ryutaro Utsumi utsumi{at}nara.kindai.ac.jp A constitutively active mutant of histidine kinase sensor EvgS was found to confer multi-drug resistance (MDR) to an acrA -deficient Escherichia coli , indicating the relationship between the two-component system EvgAS and the expression of the MDR system. The observed MDR also depended on an outer-membrane channel, TolC. Microarray and S1 mapping assays indicated that, in the presence of this constitutive mutant EvgS, the level of transcription increased for some MDR genes, including the drug efflux genes emrKY , yhiUV , acrAB , mdfA and tolC . Transcription in vitro of emrK increased by the addition of phosphorylated EvgA. Transcription activation of tolC by the activated EvgS was, however, dependent on both EvgAS and PhoPQ (Mg 2+ -responsive two-component system), in agreement with the presence of the binding site (PhoP box) for the regulator PhoP in the tolC promoter region. Transcription in vitro of yhiUV also appears to require an as-yet-unidentified additional transcriptional factor besides EvgA. Taken together we propose that the expression of the MDR system is under a complex regulatory network, including the phosphorylated EvgA serving as the master regulator. Abbreviations: MDR, multi-drug resistance