The RcsAB Box

CHARACTERIZATION OF A NEW OPERATOR ESSENTIAL FOR THE REGULATION OF EXOPOLYSACCHARIDE BIOSYNTHESIS IN ENTERIC BACTERIA*

  1. Frank Bernhard
  1. From the Freie Universität Berlin, Institut für Kristallographie, Takustrasse 6, D-14195 Berlin, Germany

Abstract

The interaction of the two transcriptional regulators RcsA and RcsB with a specific operator is a common mechanism in the activation of capsule biosynthesis in enteric bacteria. We describe RcsAB binding sites in the wza promoter of the operon for colanic acid biosynthesis in Escherichia coliK-12, in the galF promoter of the operon for K2 antigen biosynthesis in Klebsiella pneumoniae, and in thetviA (vipR) promoter of the operon for Vi antigen biosynthesis in Salmonella typhi. We further show the interaction of RcsAB with the rcsA promoters of various species, indicating that rcsA autoregulation also depends on the presence of both proteins. The compilation of all identified RcsAB binding sites revealed the conserved core sequence TaAGaatatTCctA, which we propose to be termed RcsAB box. The RcsAB box is also part ofBordetella pertussis BvgA binding sites and may represent a more distributed recognition motif within the LuxR superfamily of transcriptional regulators. The RcsAB box is essential for the induction of Rcs-regulated promoters. Site-specific mutations of conserved nucleotides in the RcsAB boxes of the E. coli wzaand rcsA promoters resulted in an exopolysaccharide-negative phenotype and in the reduction of reporter gene expression.

  • Abbreviations:
    EPS
    exopolysaccharide
    bp
    base pair(s)
    kb
    kilobase pair(s)
    ORF
    open reading frame
    PCR
    polymerase chain reaction
    EMSA
    electrophoretic mobility shift assay
    SPR
    surface plasmon resonance
    • Received October 1, 1999.
    • Revision received December 2, 1999.
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