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Journal of Bacteriology, October 2003, p. 5735-5746, Vol. 185, No. 19
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.19.5735-5746.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Genome-Wide Analyses Revealing a Signaling Network of the RcsC-YojN-RcsB Phosphorelay System in Escherichia coli

Daisuke Hagiwara,1 Masahito Sugiura,1 Taku Oshima,2 Hirotada Mori,2 Hirofumi Aiba,1 Takafumi Yamashino,1 and Takeshi Mizuno1*

Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya 464-8601,1 Research and Education Center for Genetic Information, Nara Institute of Science and Technology, Ikoma 630-0101, Japan2

Received 12 May 2003/ Accepted 10 July 2003

In Escherichia coli, capsular colanic acid polysaccharide synthesis is regulated through the multistep RcsC->YojN->RcsB phosphorelay. By monitoring a hallmarked cps::lacZ reporter gene, we first searched for physiological stimuli that propagate the Rcs signaling system. The expression of cps::lacZ was activated when cells were grown at a low temperature (20°C) in the presence of glucose as a carbon source and in the presence of a relatively high concentration of external zinc (1 mM ZnCl2). In this Rcs signaling system, the rcsF gene product (a putative outer membrane-located lipoprotein) was also an essential signaling component. Based on the defined signaling pathway and physiological stimuli for the Rcs signaling system, we conducted genome-wide analyses with microarrays to clarify the Rcs transcriptome (i.e., Rcs regulon). Thirty-two genes were identified as putative Rcs regulon members; these genes included 15 new genes in addition to 17 of the previously described cps genes. Using a set of 37 two-component system mutants, we performed alternative genome-wide analyses. The results showed that the propagation of the zinc-responsive Rcs signaling system was largely dependent on another two-component system, PhoQ/P. Considering the fact that the PhoQ/P signaling system responds to external magnesium, we obtained evidence which supports the view that there is a signaling network that connects the Rcs system with the PhoQ/P system, which coordinately regulates extracellular polysaccharide synthesis in response to the external concentrations of divalent cations.

* Corresponding author. Mailing address: Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan. Phone: 81 52 7894089. Fax: 81 52 7894091. E-mail: tmizuno{at}agr.nagoya-u.ac.jp.

Journal of Bacteriology, October 2003, p. 5735-5746, Vol. 185, No. 19
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.19.5735-5746.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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