Ribosomal Protein S1 Specifically Binds to the 5′ Untranslated Region of the Pseudomonas aeruginosa Stationary-Phase Sigma Factor rpoS mRNA in the Logarithmic Phase of Growth

ABSTRACT The rpoS gene encodes the stationary-phase sigma factor (RpoS or σs), which was identified in several gram-negative bacteria as a central regulator controlling the expression of genes involved in cell survival in response to cessation of growth (stationary phase) and providing cross-protection against various stresses. In Pseudomonas aeruginosa, the levels of σs increase dramatically at the onset of the stationary phase and are regulated at the transcriptional and posttranscriptional levels. The P. aeruginosa rpoS gene is transcribed as a monocistronic rpoS mRNA transcript comprised of an unusually long 373-bp 5′ untranslated region (5′ UTR). In this study, the 5′ UTR and total protein extracts from P. aeruginosa logarithmic and stationary phases of growth were used in order to investigate the protein-RNA interactions that may modulate the translational process. It was observed that a 69-kDa protein, which corresponded to ribosomal protein S1, preferentially binds the 5′ UTR of the rpoS mRNA in the logarithmic phase and not in the stationary phase. This is the first report of a protein-rpoS mRNA 5′ UTR interaction in P. aeruginosa, and the possible involvement of protein S1 in translation regulation of rpoS is discussed.

[1]  I. Boni,et al.  A key role for the mRNA leader structure in translational control of ribosomal protein S1 synthesis in gamma-proteobacteria. , 2003, Nucleic acids research.

[2]  B. Rolfe,et al.  Proteomic Analysis of Wild-Type Sinorhizobium meliloti Responses to N-Acyl Homoserine Lactone Quorum-Sensing Signals and the Transition to Stationary Phase , 2003, Journal of bacteriology.

[3]  V. Venturi,et al.  Role of GacA, LasI, RhlI, Ppk, PsrA, Vfr and ClpXP in the regulation of the stationary-phase sigma factor rpoS/RpoS in Pseudomonas , 2003, Archives of Microbiology.

[4]  David J. Worhunsky,et al.  Interactions of the Non-coding RNA DsrA and RpoS mRNA with the 30 S Ribosomal Subunit* , 2003, The Journal of Biological Chemistry.

[5]  N. Majdalani,et al.  Regulation and mode of action of the second small RNA activator of RpoS translation, RprA , 2002, Molecular microbiology.

[6]  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.

[7]  B. Iglewski,et al.  Role of the Pseudomonas aeruginosa las and rhl quorum-sensing systems in rhlI regulation. , 2002, FEMS microbiology letters.

[8]  Vittorio Venturi,et al.  TetR Family Member PsrA Directly Binds the Pseudomonas rpoS and psrA Promoters , 2002, Journal of bacteriology.

[9]  J. Frank,et al.  Visualization of protein S1 within the 30S ribosomal subunit and its interaction with messenger RNA , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Stanley N. Cohen,et al.  Escherichia coli Poly(A)-binding Proteins That Interact with Components of Degradosomes or Impede RNA Decay Mediated by Polynucleotide Phosphorylase and RNase E* , 2001, The Journal of Biological Chemistry.

[11]  N. V. Tzareva,et al.  Non‐canonical mechanism for translational control in bacteria: synthesis of ribosomal protein S1 , 2001, The EMBO journal.

[12]  V. Venturi,et al.  Regulation of rpoS Gene Expression in Pseudomonas: Involvement of a TetR Family Regulator , 2001, Journal of bacteriology.

[13]  F. Baralle,et al.  Splicing of constitutive upstream introns is essential for the recognition of intra-exonic suboptimal splice sites in the thrombopoietin gene. , 2001, Nucleic acids research.

[14]  R. Hengge-aronis,et al.  The Escherichia coli histone‐like protein HU regulates rpoS translation , 2001, Molecular microbiology.

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

[16]  M. Dreyfus,et al.  The Last RNA-Binding Repeat of the Escherichia coliRibosomal Protein S1 Is Specifically Involved in Autogenous Control , 2000, Journal of bacteriology.

[17]  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.

[18]  D. Hassett,et al.  Effect of rpoS Mutation on the Stress Response and Expression of Virulence Factors in Pseudomonas aeruginosa , 1999, Journal of bacteriology.

[19]  M. Bally,et al.  RpoS-dependent stress tolerance in Pseudomonas aeruginosa. , 1999, Microbiology.

[20]  C. Whistler,et al.  The Two-Component Regulators GacS and GacA Influence Accumulation of the Stationary-Phase Sigma Factor ςS and the Stress Response in Pseudomonas fluorescensPf-5 , 1998, Journal of bacteriology.

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

[22]  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.

[23]  S. Molin,et al.  Cloning, Sequencing, and Phenotypic Characterization of the rpoS Gene from Pseudomonas putida KT2440 , 1998, Journal of bacteriology.

[24]  S. Ueda,et al.  Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli: intracellular levels of four species of sigma subunit under various growth conditions , 1996, Journal of bacteriology.

[25]  R. Hengge-aronis,et al.  The RNA-binding protein HF-I, known as a host factor for phage Qbeta RNA replication, is essential for rpoS translation in Escherichia coli. , 1996, Genes & development.

[26]  A. Sarniguet,et al.  The sigma factor sigma s affects antibiotic production and biological control activity of Pseudomonas fluorescens Pf-5. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[27]  P. Seed,et al.  Activation of the Pseudomonas aeruginosa lasI gene by LasR and the Pseudomonas autoinducer PAI: an autoinduction regulatory hierarchy , 1995, Journal of bacteriology.

[28]  L. Gold,et al.  The bacteriophage T4 regB ribonuclease. Stimulation of the purified enzyme by ribosomal protein S1. , 1994, The Journal of biological chemistry.

[29]  D. Wozniak,et al.  Transcriptional analysis of the Pseudomonas aeruginosa genes algR, algB, and algD reveals a hierarchy of alginate gene expression which is modulated by algT , 1994, Journal of bacteriology.

[30]  Kan Tanaka,et al.  Transcription of the principal sigma‐factor genes, rpoD and rpoS, in Pseudomonas aeruginosa is controlled according to the growth phase , 1994, Molecular microbiology.

[31]  J. Rabinowitz,et al.  The effect of Escherichia coli ribosomal protein S1 on the translational specificity of bacterial ribosomes. , 1989, The Journal of biological chemistry.

[32]  G. Ditta,et al.  Structural relationships among rhizobium meliloti symbiotic promoters , 1983, Cell.

[33]  D. Hanahan.  Studies on transformation of Escherichia coli with plasmids. , 1983, Journal of molecular biology.

[34]  D. Helinski,et al.  Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[35]  A. Ishihama.  Functional modulation of Escherichia coli RNA polymerase. , 2000, Annual review of microbiology.

[36]  R. Sparling,et al.  Regulation in the rpoS regulon of Escherichia coli. , 1998, Canadian journal of microbiology.

[37]  H. Birnboim,et al.  A rapid alkaline extraction method for the isolation of plasmid DNA. , 1983, Methods in enzymology.

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