One ligand, two regulators and three binding sites: How KDPG controls primary carbon metabolism in Pseudomonas

PLoS Genet. 2017 Jun 28;13(6):e1006839. doi: 10.1371/journal.pgen.1006839. eCollection 2017 Jun.

Abstract

Effective regulation of primary carbon metabolism is critically important for bacteria to successfully adapt to different environments. We have identified an uncharacterised transcriptional regulator; RccR, that controls this process in response to carbon source availability. Disruption of rccR in the plant-associated microbe Pseudomonas fluorescens inhibits growth in defined media, and compromises its ability to colonise the wheat rhizosphere. Structurally, RccR is almost identical to the Entner-Doudoroff (ED) pathway regulator HexR, and both proteins are controlled by the same ED-intermediate; 2-keto-3-deoxy-6-phosphogluconate (KDPG). Despite these similarities, HexR and RccR control entirely different aspects of primary metabolism, with RccR regulating pyruvate metabolism (aceEF), the glyoxylate shunt (aceA, glcB, pntAA) and gluconeogenesis (pckA, gap). RccR displays complex and unusual regulatory behaviour; switching repression between the pyruvate metabolism and glyoxylate shunt/gluconeogenesis loci depending on the available carbon source. This regulatory complexity is enabled by two distinct pseudo-palindromic binding sites, differing only in the length of their linker regions, with KDPG binding increasing affinity for the 28 bp aceA binding site but decreasing affinity for the 15 bp aceE site. Thus, RccR is able to simultaneously suppress and activate gene expression in response to carbon source availability. Together, the RccR and HexR regulators enable the rapid coordination of multiple aspects of primary carbon metabolism, in response to levels of a single key intermediate.

MeSH terms

  • Bacterial Proteins / genetics*
  • Binding Sites
  • Carbon / metabolism
  • Gene Expression Regulation, Bacterial
  • Gluconates / metabolism*
  • Gluconeogenesis / genetics
  • Glucose / metabolism
  • Glyoxylates / metabolism
  • Ligands
  • Metabolic Networks and Pathways / genetics
  • Pseudomonas fluorescens / genetics*
  • Pseudomonas fluorescens / metabolism
  • Pyruvic Acid / metabolism
  • Transcription Factors / genetics*

Substances

  • Bacterial Proteins
  • Gluconates
  • Glyoxylates
  • Ligands
  • Transcription Factors
  • 2-keto-3-deoxy-6-phosphogluconate
  • Carbon
  • Pyruvic Acid
  • Glucose
  • glyoxylic acid

Grants and funding

This work was supported by Biotechnology and Biological Sciences Research Council (BBSRC) Institute Strategic Program grant BB/J004553/1, and University of East Anglia start-up funding to JGM. RC was supported by a 2 years postdoctoral fellowship from Istituto Pasteur Fondazione Cenci-Bolognetti, Roma (IT). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.