RESUMO
The Rcs phosphorelay signal transduction system of Escherichia coli controls genes for capsule production and many other envelope-related functions and is implicated in biofilm formation. The outer-membrane lipoprotein RcsF is an essential component of the Rcs system. Mislocalization of RcsF to the periplasm or the cytoplasmic membrane leads to high activation of the Rcs system, suggesting that RcsF functions by interacting with the cytoplasmic membrane component(s) of the system in activating the system. This is consistent with the result reported by Cho et al. (Cell159, 1652-1664, 2014) showing that RcsF interacts with the periplasmic domain (YrfFperi) of the inner-membrane protein YrfF (IgaA in Salmonella enterica serovar Typhimurium), which is a negative regulator of the Rcs system. In this study we show that RcsF also interacts with the periplasmic domain of the innermembrane-localized histidine kinase RcsC (RcsCperi). RcsCperi, which was secreted to the periplasm by fusion to maltose-binding protein, titrated RcsF's activating effect. A bimolecular fluorescence complementation experiment showed interaction of RcsF with RcsCperi, as well as with YrfFperi. We conclude that RcsF interacts with the periplasmically exposed region of RcsC, as well as with that of YrfF.