Abstract

Bacteria adapt to versatile environments by modulating gene expression through a set of stress response regulators, alternative Sigma factors, or two-component systems. Among the central processes that must be finely tuned is membrane homeostasis, including synthesis of phospholipids (PL). However, few genetic regulations of this process have been reported. We have previously shown that the gene coding the first step of PL synthesis is regulated by σE and ppGpp, and that the BasRS (PmrAB) two component system controls the expression of the DgkA PL recycling enzyme. The gene coding for phosphatidylserine decarboxylase, the last step in phosphatidylethanolamine synthesis is another gene in the PL synthesis pathway susceptible of stress response regulation. Indeed, psd appears in transcriptome studies of the σE envelope stress Sigma factor and of the CpxAR two component system. Interestingly, this gene is presumably in operon with mscM coding for a miniconductance mechanosensitive channel. In this study, we dissected the promoter region of the psd-mscM operon and studied its regulation by σE and CpxR. By artificial activation of σE and CpxRA stress response pathways, using GFP transcriptional fusion and western-blot analysis of Psd and MscM enzyme production, we showed that the operon is under the control of two distinct promoters. One is activated by σE, the second is activated by CpxRA and also responsible for basal expression of the operon. The fact that the phosphatidylethanolamine synthesis pathway is controlled by envelope stress responses at both its first and last steps might be important for adaptation of the membrane to envelope perturbations.

Highlights

  • Phospholipids (PL) are major structural and functional components of biomembranes and play a dynamic role in many regulatory processes

  • We have previously shown that plsB, coding for the first acylation step in PL synthesis, is activated by the envelope stress response σE factor, and that dgkA, coding for a diacylglycerol kinase involved in PL recycling, is controlled by the two-component system BasRS (Wahl et al, 2011)

  • The goal of this work was to decipher the genetic control of psd-mscM operon and its regulation by the two envelope stress responses mediated by the alternative Sigma factor σE and by the two-component system CpxRA

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Summary

Introduction

Phospholipids (PL) are major structural and functional components of biomembranes and play a dynamic role in many regulatory processes. The biochemistry of the PL biosynthesis pathway is well described in bacteria (Parsons and Rock, 2013). It begins by two successive acylations of the positions 1 and 2 of a glycerol 3-phosphate backbone to give phosphatidic acid. The third psd Regulation by Envelope Stress Responses position of the future polar head group is activated with a CDP nucleotide. Two separate pathways give rise in Escherichia coli to the zwitterionic phosphatidylethanolamine (PE), or to the anionic phospholipids phosphatidylglycerol (PG), and cardiolipin (CL). Bacterial species display specific membrane compositions, and the less frequent phosphatidylcholine, phosphatidylinositol, and a variety of other membrane lipids can be found in addition to the common PE, PG, and CL lipids (Sohlenkamp and Geiger, 2016)

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