1887

Abstract

EIIA is a member of a truncated phosphotransferase (PTS) system that serves regulatory functions and exists in many in addition to the sugar transport PTS. In , EIIA regulates K homeostasis through interaction with the K transporter TrkA and sensor kinase KdpD. In the H16, EIIA influences formation of the industrially important bioplastic poly(3-hydroxybutyrate) (PHB). PHB accumulation is controlled by the stringent response and induced under conditions of nitrogen deprivation. Knockout of EIIA increases the PHB content. In contrast, absence of enzyme I or HPr, which deliver phosphoryl groups to EIIA, has the opposite effect. To clarify the role of EIIA in PHB formation, we screened for interacting proteins that co-purify with Strep-tagged EIIA from cells. This approach identified the bifunctional ppGpp synthase/hydrolase SpoT1, a key enzyme of the stringent response. Two-hybrid and far-Western analyses confirmed the interaction and indicated that only non-phosphorylated EIIA interacts with SpoT1. Interestingly, this interaction does not occur between the corresponding proteins of . Vice versa, interaction of EIIA with KdpD appears to be absent in , although EIIA can perfectly substitute its homologue in in regulation of KdpD activity. Thus, interaction with KdpD might be an evolutionary ‘ancient’ task of EIIA that was subsequently replaced by interaction with SpoT1 in . In conclusion, EIIA might integrate information about nutritional status, as reflected by its phosphorylation state, into the stringent response, thereby controlling cellular PHB content in .

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2014-04-01
2020-01-19
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