A two-component phosphotransfer network involving ArcB, ArcA, and RssB coordinates synthesis and proteolysis of sigmaS (RpoS) in E. coli.

The general stress sigma factor sigma(S) (RpoS) in Escherichia coli is controlled at the levels of transcription, translation, and proteolysis. Here we demonstrate that the phosphorylated response regulator ArcA is a direct repressor of rpoS transcription that binds to two sites flanking the major rpoS promoter, with the upstream site overlapping an activating cAMP-CRP-binding site. The histidine sensor kinase ArcB not only phosphorylates ArcA, but also the sigma(S) proteolytic targeting factor RssB, and thereby stimulates sigma(S) proteolysis. Thus, ArcB/ArcA/RssB constitute a branched "three-component system", which coordinates rpoS transcription and sigma(S) proteolysis and thereby maintains low sigma(S) levels in rapidly growing cells. We suggest that the redox state of the quinones, which controls autophosphorylation of ArcB, not only monitors oxygen but also energy supply, and we show that the ArcB/ArcA/RssB system is involved in sigma(S) induction during entry into starvation conditions. Moreover, this induction is enhanced by a positive feedback that involves sigma(S)-dependent induction of ArcA, which further reduces sigma(S) proteolysis, probably by competing with RssB for residual phosphorylation by ArcB.

[1]  Svetlana Alexeeva,et al.  Requirement of ArcA for Redox Regulation in Escherichia coli under Microaerobic but Not Anaerobic or Aerobic Conditions , 2003, Journal of bacteriology.

[2]  R. Hengge-aronis,et al.  Regulation of RssB‐dependent proteolysis in Escherichia coli: a role for acetyl phosphate in a response regulator‐controlled process , 1998, Molecular microbiology.

[3]  R. Hengge-aronis,et al.  Role of the response regulator RssB in σS recognition and initiation of σS proteolysis in Escherichia coli , 2001 .

[4]  Rapid confirmation of single copy lambda prophage integration by PCR. , 1994, Nucleic acids research.

[5]  A. S. Lynch,et al.  The ArcB Sensor Kinase of Escherichia coli: Genetic Exploration of the Transmembrane Region , 2000, Journal of bacteriology.

[6]  J. Stock,et al.  Acetyl phosphate and the activation of two-component response regulators. , 1994, The Journal of biological chemistry.

[7]  F. Neidhardt,et al.  Escherichia Coli and Salmonella: Typhimurium Cellular and Molecular Biology , 1987 .

[8]  Xueqiao Liu,et al.  Probing the ArcA-P Modulon of Escherichia coli by Whole Genome Transcriptional Analysis and Sequence Recognition Profiling* , 2004, Journal of Biological Chemistry.

[9]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[10]  E C Lin,et al.  A weight matrix for binding recognition by the redox‐response regulator ArcA‐P of Escherichia coli , 1999, Molecular microbiology.

[11]  Yong Sun Lee,et al.  Multimerization of Phosphorylated and Non-phosphorylated ArcA Is Necessary for the Response Regulator Function of the Arc Two-component Signal Transduction System* , 2001, The Journal of Biological Chemistry.

[12]  R. Hengge-aronis,et al.  The cellular level of the recognition factor RssB is rate‐limiting for σS proteolysis: implications for RssB regulation and signal transduction in σS turnover in Escherichia coli , 2002 .

[13]  D. Georgellis,et al.  Identification of a quinone-sensitive redox switch in the ArcB sensor kinase. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[14]  G. Storz,et al.  The response regulator RssB controls stability of the sigma(S) subunit of RNA polymerase in Escherichia coli. , 1996, The EMBO journal.

[15]  T. Mizuno,et al.  Phosphotransfer circuitry of the putative multi‐signal transducer, ArcB, of Escherichia coli: in vitro studies with mutants , 1995, Molecular microbiology.

[16]  N. Majdalani,et al.  Small non‐coding RNAs, co‐ordinators of adaptation processes in Escherichia coli: the RpoS paradigm , 2003, Molecular microbiology.

[17]  R. Hengge-aronis,et al.  Signal Transduction and Regulatory Mechanisms Involved in Control of the σS (RpoS) Subunit of RNA Polymerase , 2002, Microbiology and Molecular Biology Reviews.

[18]  Victor Sourjik,et al.  Receptor clustering and signal processing in E. coli chemotaxis. , 2004, Trends in microbiology.

[19]  Ka-Yiu San,et al.  Effect of oxygen on the Escherichia coli ArcA and FNR regulation systems and metabolic responses. , 2005, Biotechnology and bioengineering.

[20]  V. Wendisch,et al.  Genome-Wide Analysis of the General Stress Response Network in Escherichia coli: σS-Dependent Genes, Promoters, and Sigma Factor Selectivity , 2005, Journal of bacteriology.

[21]  R. Hengge-aronis,et al.  The response regulator RssB, a recognition factor for σS proteolysis in Escherichia coli, can act like an anti‐σS factor , 2000 .

[22]  P. Dersch,et al.  RovA is autoregulated and antagonizes H‐NS‐mediated silencing of invasin and rovA expression in Yersinia pseudotuberculosis , 2004, Molecular microbiology.

[23]  D. Georgellis,et al.  Requirement of the Receiver and Phosphotransfer Domains of ArcB for Efficient Dephosphorylation of Phosphorylated ArcA In Vivo , 2005, Journal of bacteriology.

[24]  R. Kolter,et al.  Stationary-phase-inducible "gearbox" promoters: differential effects of katF mutations and role of sigma 70 , 1991, Journal of bacteriology.

[25]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .

[26]  R. Hengge-aronis,et al.  Recent insights into the general stress response regulatory network in Escherichia coli. , 2002, Journal of molecular microbiology and biotechnology.

[27]  S. Gottesman,et al.  The RssB response regulator directly targets sigma(S) for degradation by ClpXP. , 2001, Genes & development.

[28]  R. Hengge-aronis,et al.  Regulation of RpoS proteolysis in Escherichia coli: the response regulator RssB is a recognition factor that interacts with the turnover element in RpoS. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[29]  R. Hengge-aronis,et al.  Identification of transcriptional start sites and the role of ppGpp in the expression of rpoS, the structural gene for the sigma S subunit of RNA polymerase in Escherichia coli , 1995, Journal of bacteriology.

[30]  R. Utsumi,et al.  Functional Characterization in Vitro of All Two-component Signal Transduction Systems from Escherichia coli* , 2005, Journal of Biological Chemistry.

[31]  G. W. Hatfield,et al.  Global Gene Expression Profiling in Escherichia coli K12 , 2003, Journal of Biological Chemistry.

[32]  R. Simons,et al.  Improved single and multicopy lac-based cloning vectors for protein and operon fusions. , 1987, Gene.

[33]  A. S. Lynch,et al.  Transcriptional control mediated by the ArcA two-component response regulator protein of Escherichia coli: characterization of DNA binding at target promoters , 1996, Journal of bacteriology.

[34]  Pierre Baldi,et al.  Global gene expression profiling in Escherichia coli K12: effects of oxygen availability and ArcA. , 2005, The Journal of biological chemistry.

[35]  A. Emili,et al.  Interaction network containing conserved and essential protein complexes in Escherichia coli , 2005, Nature.

[36]  U. Jenal,et al.  Regulation by proteolysis in bacterial cells. , 2003, Current opinion in microbiology.

[37]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[38]  M. Casadaban,et al.  Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. , 1976, Journal of molecular biology.

[39]  A. S. Lynch,et al.  In vitro phosphorylation study of the arc two-component signal transduction system of Escherichia coli , 1997, Journal of bacteriology.

[40]  D. Touati,et al.  Anaerobic activation of arcA transcription in Escherichia coli: roles of Fnr and ArcA , 1994, Molecular microbiology.

[41]  S. Ryu,et al.  Expression of ptsG Encoding the Major Glucose Transporter Is Regulated by ArcA in Escherichia coli* , 2004, Journal of Biological Chemistry.

[42]  E. Lin,et al.  Quinones as the Redox Signal for the Arc Two-Component System of Bacteria , 2001, Science.

[43]  E. Lin,et al.  Redox Signal Transduction by the ArcB Sensor Kinase of Haemophilus influenzae Lacking the PAS Domain , 2001, Journal of bacteriology.

[44]  E. Lin,et al.  Signal Decay through a Reverse Phosphorelay in the Arc Two-component Signal Transduction System* , 1998, The Journal of Biological Chemistry.

[45]  R. Hengge-aronis,et al.  The cellular concentration of the sigma S subunit of RNA polymerase in Escherichia coli is controlled at the levels of transcription, translation, and protein stability. , 1994, Genes & development.