Amino Acids

, Volume 40, Issue 2, pp 421–430 | Cite as

Activation of the AtoSC two-component system in the absence of the AtoC N-terminal receiver domain in E. coli

  • Evaggelos C. Theodorou
  • Marina C. Theodorou
  • Margarita N. Samali
  • Dimitrios A. Kyriakidis
Original Article
First Online:
Received:
Accepted:

Abstract

The AtoSC two-component system in E. coli consists of the AtoS sensor kinase and the AtoC response regulator. It regulates positively the transcriptional activation of atoDAEB operon, encoding enzymes involved in short-chain fatty acid catabolism upon acetoacetate-mediated induction. AtoSC acting on atoDAEB operon, regulates the biosynthesis and the intracellular distribution of short-chain poly-(R)-3-hydroxybutyrate (cPHB). A phosphorylation-incompetent AtoC form was constructed lacking its N-terminal receiver domain, trAtoC, to study the effects of AtoC domains on cPHB biosynthesis and atoDAEB operon regulation. Both cPHB biosynthesis and atoDAEB gene expression were regulated positively by trAtoC in the absence of any inducer in E. coli of both atoSC + and ΔatoSC genotypes. The presence of acetoacetate or spermidine further promoted these trAtoC actions. Competitive regulatory functions between the full length AtoC and trAtoC were observed referring to atoDAEB and cPHB targets as well as growth of trAtoC-overproducing atoSC + cells on butyrate as the sole carbon source. trAtoC in contrast to the wild-type AtoC presented different modes of cPHB and atoDAEB regulation in the presence of compounds involved in fatty acid metabolism including CoA-SH, acetyl-CoA, sodium acetate or 3-hydroxybutyryl-CoA. These data provide evidence for a role of the AtoC N-terminal receiver domain in regulating the biological activities of AtoSC as well as additional mechanisms of interactions between the AtoSC constituents including their established inducers or new effectors towards the accomplishment of the AtoSC TCS signal transduction.

Keywords

AtoSC Truncated AtoC atoDAEB Two-component system Poly-(R)-3-hydroxybutyrate 

Abbreviations

atoSC+

Genetic locus encoding the AtoS and AtoC proteins

AcAc

Acetoacetate

cPHB

Complexed poly-(R)-3-hydroxybutyrate

C48/80

Compound C48/80

HK

Histidine kinase

sPHB

Storage polyhydroxybutyrate

RR

Response regulator

TCS

Two-component system

trAtoC

N-terminal domain-truncated AtoC

URR

Upstream regulatory region

σ54-RNAP

σ54-RNA polymerase

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Notes

Acknowledgments

This work was supported by doctoral research scholarships to E.C.T. by the “State Scholarships Foundation” and “Onassis Foundation”, by “PYTHAGORAS post doctoral grant” to M.C.T. of the Greek Ministry of Education and a grant ‘‘BIOPRODUCTION’’ of the 6th FP of EC. We thank Dr. Hirofumi Aiba (Nagoya University, Japan) for E. coli K-12 strains BW25113 and BW28878.

Conflict of interest statement

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Evaggelos C. Theodorou
    • 1
  • Marina C. Theodorou
    • 1
  • Margarita N. Samali
    • 1
  • Dimitrios A. Kyriakidis
    • 1
    • 2
  1. 1.Laboratory of Biochemistry, Department of ChemistryAristotle University of ThessalonikiThessalonikiGreece
  2. 2.National Hellenic Research FoundationAthensGreece

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