1887

Abstract

Gene encodes peptidyl-tRNA hydrolase (Pth), an enzyme that cleaves peptidyl-tRNAs released abortively from ribosomes during protein synthesis. In the chromosome, is flanked by and , two genes of unknown function. Pth is essential for cell viability, especially under conditions leading to overproduction of peptidyl-tRNA. In an attempt to unveil the elements that affect expression, the transcriptional features of the region were investigated. Northern blot experiments showed that both and , the 3′-proximal gene, are cotranscribed in a bicistronic transcript. However, transcripts containing each of the individual messages were also detected. Accordingly, two transcriptional promoters were identified by primer extension experiments: one located upstream of , which presumably gives rise to both the mono and bicistronic transcripts, and the other, preceding , which generates its monocistronic message. Deletion analysis indicates that transcript stability depends on integrity. Also, a defect in RNase E activity resulted in Pth overproduction. It is proposed that RNase E processing within F in the bicistronic message limits expression.

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2002-11-01
2024-11-14
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References

  1. Aiba H, Adhya S., de Crombrugghe B. 1981; Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem 256:11905–11910
    [Google Scholar]
  2. Artsimovitch I, Kahmeyer-Gabbe M., Howe M. M. 1996; Distortion in the spacer region of Pm during activation of middle transcription of phage Mu. Proc Natl Acad Sci USA 93:9408–9413
    [Google Scholar]
  3. Atherly A. G., Menninger J. R. 1972; Mutant E. coli strain with temperature sensitive peptidyl-transfer RNA hydrolase. Nature 240:245–246
    [Google Scholar]
  4. Auble D. T., deHaseth P. L. 1988; Promoter recognition by Escherichia coli RNA polymerase. Influence of DNA structure in the spacer separating the −10 and −35 regions. J Mol Biol 202:471–482
    [Google Scholar]
  5. Bardwell J. C, Régnier P, Chen S. M, Nakamura Y, Grunberg-Manago M., Court D. L. 1989; Autoregulation of RNase III operon by mRNA processing. EMBO J 8:3401–3407
    [Google Scholar]
  6. Cruz-Vera L. R, Toledo I, Hernández-Sánchez J., Guarneros G. 2000; Molecular basis for the temperature sensivity of Escherichia coli pth (Ts). J Bacteriol 182:1523–1528
    [Google Scholar]
  7. De la Vega F. M, Galindo J. M, Gold I. G., Guarneros G. 1996; Microbial genes homologous to the peptidyl-tRNA hydrolase-encoding gene of Escherichia coli . Gene 169:97–100
    [Google Scholar]
  8. Dinçbas V, Heurgué-Hamard V, Buckingham R. H, Karimi R., Ehrenberg M. 1999; Shutdown in protein synthesis due to the expression of mini-genes in bacteria. J Mol Biol 291:745–759
    [Google Scholar]
  9. Dutka S, Meinnel T, Lazennec C, Mechulam Y., Blanquet S. 1993; Role of the 1–72 base pair in tRNAs for the activity of Escherichia coli peptidyl-tRNA hydrolase. Nucleic Acids Res 21:4025–4030
    [Google Scholar]
  10. Fleischmann R. D, Adams M. D, White O. 37 other authors 1995; Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science 269:496–512
    [Google Scholar]
  11. Fromant M, Plateau P, Schmitt E, Mechulam Y., Blanquet S. 1999; Receptor site for the 5′-phosphate of elongator tRNAs governs substrate selection by peptidyl-tRNA hydrolase. Biochemistry 38:4982–4987
    [Google Scholar]
  12. Galindo J. M, Guarneros G., De La Vega F. M. 1994; Open reading frames flanking the peptidyl-tRNA hydrolase-encoding gene of Escherichia coli . Gene 151:153–156
    [Google Scholar]
  13. Garcı́a-Mena J, Das A, Sánchez-Trujillo A, Portier C., Montañez C. 1999; A novel mutation in the KH domain of polynucleotide phosphorylase affects autoregulation and mRNA decay in Escherichia coli . Mol Microbiol 33:235–248
    [Google Scholar]
  14. Garcı́a-Villegas R, De La Vega F. M, Galindo J. M, Segura M, Buckingham R. H., Guarneros G. 1991; Peptidyl-tRNA hydrolase is involved in λ inhibition of host protein synthesis. EMBO J 10:3549–3555
    [Google Scholar]
  15. Guzmán G., Guarneros G. 1989; Phage genetic sites involved in lambda growth inhibition by Escherichia coli rap mutants. Genetics 21:401–409
    [Google Scholar]
  16. Hajnsdorf E, Carpousis A. J., Régnier P. 1994; Nucleolytic inactivation and degradation of RNAse III processed pnp message encoding polynucleotide phosphorylase of Escherichia coli . J Mol Biol 239:439–454
    [Google Scholar]
  17. Hawley D. K., McClure W. R. 1983; Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res 11:2237–2255
    [Google Scholar]
  18. Hernández J, Ontiveros C, Valadez J. G, Buckingham R. H., Guarneros G. 1997; Regulation of protein synthesis by minigene expression. Biochimie 79:527–531
    [Google Scholar]
  19. Hernández-Sánchez J, Valadez J. G, Vega-Herrera J, Ontiveros C., Guarneros G. 1998; Lambda bar minigene-mediated inhibition of protein synthesis involves accumulation of peptidyl-tRNA and starvation for tRNA. EMBO J 17:3758–3765
    [Google Scholar]
  20. Heurgué-Hamard V, Karimi R, Mora L, MacDougall J, Leboeuf C, Grentzmann G, Ehrenberg M., Buckingham R. H. 1998; Ribosome release factor RF4 and termination factor RF3 are involved in dissociation of peptidyl-tRNA from the ribosome. EMBO J 17:808–816
    [Google Scholar]
  21. Heurgué-Hamard V, Dinçbas V, Buckingham R. H., Ehrenberg M. 2000; Origins of minigene-dependent growth inhibition in bacterial cells. EMBO J 19:2701–2709
    [Google Scholar]
  22. Higgins C. F, Causton H. C, Dance G. S. C., Mudd H. A. 1993; RNA degradation. In Control of Messenger RNA Stability pp 13–30 Edited by Belasco J., Brawerman G. San Diego, CA: Academic Press;
    [Google Scholar]
  23. Jain C., Belasco J. G. 1995; RNase E autoregulates its synthesis by controlling the degradation rate of its own mRNA in Escherichia coli : unusual sensitivity of the rne transcript to RNase E activity. Genes Dev 9:84–96
    [Google Scholar]
  24. Kohara Y, Akiyama K., Isono K. 1987; The physical map of the whole E. coli chromosome; application of a new strategy for rapid analysis and sorting of large genomic library. Cell 50:495–508
    [Google Scholar]
  25. Lesnik E. A, Sampath R, Levene H. B, Henderson T. J, McNeil J. A., Ecker D. J. 2001; Prediction of rho-independent transcriptional terminators in Escherichia coli . Nucleic Acids Res 29:3583–3594
    [Google Scholar]
  26. Manson P. J., Williams J. G. 1985; Hybridisation in the analysis of recombination DNA. In Nucleic Acid Hybridisation, a Practical Approach pp 213–276 Edited by Hames B. D., Higgins S. J. Oxford: IRL Press;
    [Google Scholar]
  27. Mateos L. M, Pisabarro A, Pátek M, Malumbres M, Guerrero C, Eikmanns B. J, Sahm H., Martı́n J. F. 1994; Transcriptional analysis and regulatory signals of the hom–thrB cluster of Brevibacterium lactofermentum . J Bacteriol 176:7362–7371
    [Google Scholar]
  28. McClelland M, Sanderson K. E, Spieth J. 23 other authors 2001; Complete genome sequence of Salmonella enterica serovar typhimurium LT2. Nature 413:852–856
    [Google Scholar]
  29. McDowall K. J, Hernández R. G, Lin-Chao S., Cohen S. N. 1993; The ams -1 and rne -3071 temperature-sensitive mutations in the ams gene are in close proximity to each other and cause substitutions within a domain that resembles a product of the Escherichia coli mre locus. J Bacteriol 175:4245–4249
    [Google Scholar]
  30. Menez J, Heurgué-Hamard V., Buckingham R. H. 2000; Sequestration of specific tRNA species cognate to the last sense codon of an overproduced gratuitous protein. Nucleic Acids Res 28:4725–4732
    [Google Scholar]
  31. Nickerson C. A., Achberger E. C. 1995; Role of curved DNA in binding of Escherichia coli RNA polymerase to promoters. J Bacteriol 177:5756–5761
    [Google Scholar]
  32. Ontiveros C, Valadez J. G, Hernández J., Guarneros G. 1997; Inhibition of Escherichia coli protein synthesis by abortive translation of phage lambda minigenes. J Mol Biol 269:167–175
    [Google Scholar]
  33. Oshima T, Aiba H, Baba T. 52 other authors 1996; A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 12·7–28·0 min region on the linkage map. DNA Res 3:137–155
    [Google Scholar]
  34. Parkhill J, Wren B. W, Thomson N. R. 38 other authors 2001; Genome sequence of Yersinia pestis , the causative agent of plague. Nature 413:469–470
    [Google Scholar]
  35. Régnier P., Portier C. 1986; Initiation, attenuation and RNase III processing of transcripts from the Escherichia coli operon encoding ribosomal protein S15 and polynucleotide phosphorylase. J Mol Biol 187:23–32
    [Google Scholar]
  36. Rudd K. E. 1992; Alignment of E. coli DNA sequences to a revised, integrated genomic restriction map. In A Short Course in Bacterial Genetics: a Laboratory Manual and Handbook for E. coli and Related Bacteria pp 2.3–2.43 Edited by Miller J. H. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  37. Schmitt E, Mechulam Y, Fromant M, Plateau P., Blanquet S. 1997; Crystal structure at 1·2 Å resolution and active site mapping of Escherichia coli peptidyl-tRNA hydrolase. EMBO J 16:4760–4769
    [Google Scholar]
  38. Stover C. K, Pham X. Q, Erwin A. L. 28 other authors 2000; Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 406:959–964
    [Google Scholar]
  39. Wosten M. M. 1998; Eubacterial sigma-factors. FEMS Microbiol Rev 22:127–150
    [Google Scholar]
  40. Zilhao R, Régnier P., Arraiano C. M. 1995; The role of endonucleases in the expression of ribonuclease II in Escherichia coli . FEMS Microbiol Lett 130:237–244
    [Google Scholar]
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