The role of the two-component systems Cpx and Arc in protein alterations upon gentamicin treatment in Escherichia coli

Emina Ćudić,Kristin Surmann,Gianna Panasia,Sabine Hunke,Elke Hammer

doi:10.1186/s12866-017-1100-9

Emina Ćudić, Kristin Surmann + Show 3 more

Open Access

https://doi.org/10.1186/s12866-017-1100-9

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Abstract

BackgroundThe aminoglycoside antibiotic gentamicin was supposed to induce a crosstalk between the Cpx- and the Arc-two-component systems (TCS). Here, we investigated the physical interaction of the respective TCS components and compared the results with their respective gene expression and protein abundance. The findings were interpreted in relation to the global proteome profile upon gentamicin treatment.ResultsWe observed specific interaction between CpxA and ArcA upon treatment with the aminoglycoside gentamicin using Membrane-Strep-tagged protein interaction experiments (mSPINE). This interaction was neither accompanied by detectable phosphorylation of ArcA nor by activation of the Arc system via CpxA. Furthermore, no changes in absolute amounts of the Cpx- and Arc-TCS could be determined with the sensitive single reaction monitoring (SRM) in presence of gentamicin. Nevertheless, upon applying shotgun mass spectrometry analysis after treatment with gentamicin, we observed a reduction of ArcA ~ P-dependent protein synthesis and a significant Cpx-dependent alteration in the global proteome profile of E. coli.ConclusionsThis study points to the importance of the Cpx-TCS within the complex regulatory network in the E. coli response to aminoglycoside-caused stress.

Highlights

The aminoglycoside antibiotic gentamicin was supposed to induce a crosstalk between the Cpx- and the Arc-two-component systems (TCS)
Cpx and ArcA interact upon aminoglycoside treatment In order to investigate the postulated crosstalk between the Cpx- and Arc-TCS (CpxA with ArcA or ArcB with CpxR) after aminoglycoside treatment in E. coli ([8]; Fig. 1), we first analyzed the physical interactions between the sensor kinases (SK) CpxA and ArcB with the response regulators (RR) CpxR and ArcA of E. coli strain MG1655 in vivo
The SKs CpxA and ArcB were C-terminally fused to T18, whereas the RRs CpxR and ArcA were N-terminally fused to T25

Summary

Introduction

The aminoglycoside antibiotic gentamicin was supposed to induce a crosstalk between the Cpx- and the Arc-two-component systems (TCS). Treatment with aminoglycosides is commonly used to understand cellular processes that enable bacteria to resist these antibiotics. Biofilm formation and two-component system (TCS) signaling were found to increase the resistance towards aminoglycosides [2, 3]. Ćudić et al BMC Microbiology (2017) 17:197 i) integration of additional signals by an accessory protein regulated by another TCS, ii) phosphorylation of another, non-cognate RR by the SK, and iii) target promotor recognition by a non-cognate RR [11]. Such crosstalk between TCS in E. coli has been already reported between the CpxTCS and the EnvZ-OmpR-TCS [12, 13], or the quorumsensing QseBC-TCS [14]

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