Signal Transduction Cascade for Regulation of RpoS: Temperature Regulation of DsrA

ABSTRACT Many environmental parameters modulate the amount of the RpoS sigma factor in Escherichia coli. Temperature control of RpoS depends on the untranslated RNA DsrA. DsrA activates RpoS translation by pairing with the leader of the mRNA. We find that temperature affects both the rate of transcription initiation of thedsrA gene and the stability of DsrA RNA. Both are increased at low temperature (25°C) compared to 37 or 42°C. The combination of these results is 25-fold-less DsrA at 37°C and 30-fold less at 42°C than at 25°C. Using an adaptedlacZ-based reporter system, we show that temperature control of transcription initiation of dsrA requires only the minimal promoter of 36 bp. Overall, transcription responses to temperature lead to a sixfold increase in DsrA synthesis at 25°C over that at 42°C. Furthermore, two activating regions and a site for LeuO negative regulation were identified in the dsrApromoter. The activating regions also activate transcription in vitro. DsrA decays with a half-life of 23 min at 25°C and 4 min at 37 and 42°C. These results demonstrate that the dsrA promoter and the stability of DsrA RNA are the thermometers for RpoS temperature sensing. Multiple inputs to DsrA accumulation allow sensitive modulation of changes in the synthesis of the downstream targets of DsrA such as RpoS.

[1]  R. Hurme,et al.  Temperature sensing in bacterial gene regulation — what it all boils down to , 1998, Molecular microbiology.

[2]  P. Dennis,et al.  Expression of lacZ from the Promoter of theEscherichia coli spc Operon Cloned into Vectors Carrying the W205 trp-lac Fusion , 1998, Journal of bacteriology.

[3]  D. Low,et al.  An essential role for DNA adenine methylation in bacterial virulence. , 1999, Science.

[4]  H. Krisch,et al.  Specificity of Escherichia coli endoribonuclease RNase E: in vivo and in vitro analysis of mutants in a bacteriophage T4 mRNA processing site. , 1992, Genes & development.

[5]  A. Oppenheim,et al.  Enhanced activity of the bacteriophage lambda PL promoter at low temperature. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[6]  F. Repoila,et al.  Genomic polymorphism in the T‐even bacteriophages. , 1994, The EMBO journal.

[7]  C. Gualerzi,et al.  Post‐transcriptional regulation of CspA expression in Escherichia coli , 1996, Molecular microbiology.

[8]  M. Inouye,et al.  Cold-shock response and cold-shock proteins. , 1999, Current opinion in microbiology.

[9]  F. Neidhardt,et al.  Levels of major proteins of Escherichia coli during growth at different temperatures , 1979, Journal of bacteriology.

[10]  S. Gottesman,et al.  Regulation of Proteolysis of the Stationary-Phase Sigma Factor RpoS , 1998, Journal of bacteriology.

[11]  S. Cohen,et al.  Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[12]  K. Drlica,et al.  DNA switches for thermal control of gene expression. , 1999, Trends in microbiology.

[13]  G. Storz,et al.  A Small, Stable RNA Induced by Oxidative Stress: Role as a Pleiotropic Regulator and Antimutator , 1997, Cell.

[14]  A. Zhang,et al.  Hfq Is Necessary for Regulation by the Untranslated RNA DsrA , 2001, Journal of bacteriology.

[15]  K. Tilly,et al.  Temperature-regulated expression of bacterial virulence genes. , 2000, Microbes and infection.

[16]  C. Harley,et al.  Analysis of E. coli promoter sequences. , 1987, Nucleic acids research.

[17]  D. de Mendoza,et al.  Transcriptional Control of the Low-Temperature-Inducible des Gene, Encoding the Δ5 Desaturase of Bacillus subtilis , 1999, Journal of bacteriology.

[18]  R. Hengge-aronis,et al.  Posttranscriptional osmotic regulation of the sigma(s) subunit of RNA polymerase in Escherichia coli , 1996, Journal of bacteriology.

[19]  S. Gottesman,et al.  The small RNA, DsrA, is essential for the low temperature expression of RpoS during exponential growth in Escherichia coli. , 1996, The EMBO journal.

[20]  P. Tortora,et al.  Transcriptional and post‐transcriptional control of polynucleotide phosphorylase during cold acclimation in Escherichia coli , 2000, Molecular microbiology.

[21]  J. Brosius,et al.  Spacing of the -10 and -35 regions in the tac promoter. Effect on its in vivo activity. , 1985, The Journal of biological chemistry.

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

[23]  R. Simons,et al.  Regulation of the "tetCD" genes of transposon Tn10. , 1997, Journal of molecular biology.

[24]  S Falkow,et al.  Copyright © 1997, American Society for Microbiology Common Themes in Microbial Pathogenicity Revisited , 2022 .

[25]  Jürgen Böhringer,et al.  The LysR‐like regulator LeuO in Escherichia coli is involved in the translational regulation of rpoS by affecting the expression of the small regulatory DsrA‐RNA , 1997, Molecular microbiology.

[26]  A. Conter,et al.  Role of DNA supercoiling and rpoS sigma factor in the osmotic and growth phase-dependent induction of the gene osmE of Escherichia coli K12. , 1997, Journal of molecular biology.

[27]  N. Majdalani,et al.  Regulation of RpoS by a novel small RNA: the characterization of RprA. , 2001, Molecular microbiology.

[28]  A. Oppenheim,et al.  Differential mRNA stability of the cspA gene in the cold‐shock response of Escherichia coli , 1996, Molecular microbiology.

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

[30]  N. Majdalani,et al.  DsrA RNA regulates translation of RpoS message by an anti-antisense mechanism, independent of its action as an antisilencer of transcription. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[31]  A. Oppenheim,et al.  Enhanced activity of the bacteriophage λ PL promoter at low temperature , 1995 .

[32]  S. Altuvia,et al.  Alternative mRNA structures of the cIII gene of bacteriophage lambda determine the rate of its translation initiation. , 1989, Journal of molecular biology.

[33]  W. Reznikoff,et al.  Genetic fusions defining trp and lac operon regulatory elements. , 1975, Journal of molecular biology.

[34]  J. Harel,et al.  Virulence gene regulation in pathogenic Escherichia coli. , 1999, Veterinary research.

[35]  Nan Wang,et al.  CspI, the Ninth Member of the CspA Family ofEscherichia coli, Is Induced upon Cold Shock , 1999, Journal of bacteriology.

[36]  D. Jin,et al.  The rpoB mutants destabilizing initiation complexes at stringently controlled promoters behave like "stringent" RNA polymerases in Escherichia coli. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[37]  G. Storz,et al.  The OxyS regulatory RNA represses rpoS translation and binds the Hfq (HF‐I) protein , 1998, The EMBO journal.

[38]  R. Miller,et al.  One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[39]  R. Hengge-aronis,et al.  Interplay of global regulators and cell physiology in the general stress response of Escherichia coli. , 1999, Current opinion in microbiology.

[40]  Jeffrey H. Miller,et al.  A short course in bacterial genetics , 1992 .

[41]  D. Apirion,et al.  The gene for a small stable RNA (10Sa RNA) of Escherichia coli , 1989, Molecular microbiology.

[42]  J. Choih,et al.  Regulation of the , 1996 .

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

[44]  L. Enquist,et al.  Experiments With Gene Fusions , 1984 .

[45]  Y. Kyōgoku,et al.  Translational induction of heat shock transcription factor sigma32: evidence for a built-in RNA thermosensor. , 1999, Genes & development.

[46]  S. Gottesman,et al.  A small RNA acts as an antisilencer of the H-NS-silenced rcsA gene of Escherichia coli. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[47]  N. Hoe,et al.  Temperature sensing in Yersinia pestis: translation of the LcrF activator protein is thermally regulated , 1993, Journal of bacteriology.

[48]  T. Mizuno,et al.  The leuO Gene Product Has a Latent Ability To Relieve bgl Silencing inEscherichia coli , 1998, Journal of bacteriology.

[49]  R. Gourse,et al.  Identification of an UP element consensus sequence for bacterial promoters. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[50]  R. Lease,et al.  A trans-acting RNA as a control switch in Escherichia coli: DsrA modulates function by forming alternative structures. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[51]  R. Gourse,et al.  E.coli Fis protein activates ribosomal RNA transcription in vitro and in vivo. , 1990, The EMBO journal.

[52]  M. Cusick,et al.  Riboregulation in Escherichia coli: DsrA RNA acts by RNA:RNA interactions at multiple loci. , 1998, Proceedings of the National Academy of Sciences of the United States of America.