SpoT and GppA hydrolases prevent the gratuitous activation of RelA by pppGpp in Escherichia coli

Abstract

Stringent response, a conserved regulation seen in bacteria, is effected through the modified nucleotides (p)ppGpp. The metabolic cycle of these molecules is driven by the synthase activity of RelA and SpoT and the hydrolase activity of SpoT and GppA which together sets the basal (p)ppGpp pool. Growth arrest due to (p)ppGpp accumulation from basal RelA activity apparently explained the essentiality of SpoT hydrolase function. We found, pppGpp degradation was enhanced when the SpoT hydrolase activity was lowered or eliminated and when this was alleviated by inactivation of the GppA hydrolase, gratuitous synthesis of (p)ppGpp by RelA was activated, leading to growth arrest. The RelA-ribosome interaction was not mandatory for these phenotypes. Our results show, for the first time, elevated pppGpp promoted the amplification of RelA-mediated stringent response in the absence of established RelA activating signals in the cell and the SpoT and GppA hydrolases prevented this. The accumulation of pppGpp inhibited the SpoT hydrolase activity. We propose this autocatalytic activation of RelA by pppGpp is likely to be an allosteric regulation and can result in a bistable switch.