Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1986 Sep 11;14(17):6857–6870. doi: 10.1093/nar/14.17.6857

Promoter selectivity of Escherichia coli RNA polymerase: alteration by fMet-tRNAfMet.

T Nomura, N Fujita, A Ishihama
PMCID: PMC311704  PMID: 3532031

Abstract

An in vitro mixed transcription system was employed to examine the possible alteration of the promoter selectivity of Escherichia coli RNA polymerase by specific tRNAs. Transcription in vitro was inhibited by most of the tRNAs examined, although the extent of the inhibition differed with the tRNA species. The inhibition by tRNAs was due to competition with DNA for binding RNA polymerase. This inhibitory effect remained after charging of the tRNAs with amino acids. The charging of tRNAfMet with fMet, but not with Met, abolished its inhibitory effect, and instead gave a stimulatory effect on the transcription from some promoters. These observations suggest that fMet-tRNAfMet plays a specific regulatory role in the coupling of transcription to translation.

Full text

PDF
6858

Images in this article

6862Image
on p.6862
6863Image
on p.6863
6864Image
on p.6864
6865Image
on p.6865
6866Image
on p.6866
6867Image
on p.6867

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Backman K., Ptashne M., Gilbert W. Construction of plasmids carrying the cI gene of bacteriophage lambda. Proc Natl Acad Sci U S A. 1976 Nov;73(11):4174–4178. doi: 10.1073/pnas.73.11.4174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnboim H. C. A rapid alkaline extraction method for the isolation of plasmid DNA. Methods Enzymol. 1983;100:243–255. doi: 10.1016/0076-6879(83)00059-2. [DOI] [PubMed] [Google Scholar]
  3. Caillet J., Plumbridge J. A., Springer M., Vacher J., Delamarche C., Buckingham R. H., Grunberg-Manago M. Identification of clones carrying an E. coli tRNAPhe gene by suppression of phenylalanyl-tRNA synthetase thermosensitive mutants. Nucleic Acids Res. 1983 Feb 11;11(3):727–736. doi: 10.1093/nar/11.3.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cheung A. Y., Söll D. In vivo and in vitro transcription of the Escherichia coli glutaminyl-tRNA synthetase gene. J Biol Chem. 1984 Aug 10;259(15):9953–9958. [PubMed] [Google Scholar]
  5. Debenham P. G., Pongs O., Travers A. A. Formylmethionyl-tRNA alters RNA polymerase specificity. Proc Natl Acad Sci U S A. 1980 Feb;77(2):870–874. doi: 10.1073/pnas.77.2.870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fukuda R., Iwakura Y., Ishihama A. Heterogeneity of RNA polymerase in Escherichia coli. I. A new holoenzyme containing a new sigma factor. J Mol Biol. 1974 Mar;83(3):353–367. doi: 10.1016/0022-2836(74)90284-8. [DOI] [PubMed] [Google Scholar]
  7. Gonzalez N., Wiggs J., Chamberlin M. J. A simple procedure for resolution of Escherichia coli RNA polymerase holoenzyme from core polymerase. Arch Biochem Biophys. 1977 Aug;182(2):404–408. doi: 10.1016/0003-9861(77)90521-5. [DOI] [PubMed] [Google Scholar]
  8. Harvey R. J. Growth and initiation of protein synthesis in Escherichia coli in the presence of trimethoprim. J Bacteriol. 1973 Apr;114(1):309–322. doi: 10.1128/jb.114.1.309-322.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Horii T., Ogawa T., Ogawa H. Organization of the recA gene of Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jan;77(1):313–317. doi: 10.1073/pnas.77.1.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ishii S., Kuroki K., Imamoto F. tRNAMetf2 gene in the leader region of the nusA operon in Escherichia coli. Proc Natl Acad Sci U S A. 1984 Jan;81(2):409–413. doi: 10.1073/pnas.81.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. KURLAND C. G., MAALOE O. Regulation of ribosomal and transfer RNA synthesis. J Mol Biol. 1962 Mar;4:193–210. doi: 10.1016/s0022-2836(62)80051-5. [DOI] [PubMed] [Google Scholar]
  12. Kajitani M., Ishihama A. Determination of the promoter strength in the mixed transcription system. II. Promoters of ribosomal RNA, ribosomal protein S1 and recA protein operons from Escherichia coli. Nucleic Acids Res. 1983 Jun 25;11(12):3873–3888. doi: 10.1093/nar/11.12.3873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kajitani M., Ishihama A. Determination of the promoter strength in the mixed transcription system: promoters of lactose, tryptophan and ribosomal protein L10 operons from Escherichia coli. Nucleic Acids Res. 1983 Feb 11;11(3):671–686. doi: 10.1093/nar/11.3.671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kajitani M., Ishihama A. Promoter selectivity of Escherichia coli RNA polymerase. Differential stringent control of the multiple promoters from ribosomal RNA and protein operons. J Biol Chem. 1984 Feb 10;259(3):1951–1957. [PubMed] [Google Scholar]
  15. Kung H., Spears C., Weissbach H. Purification and properties of a soluble factor required for the deoxyribonucleic acid-directed in vitro synthesis of beta-galactosidase. J Biol Chem. 1975 Feb 25;250(4):1556–1562. [PubMed] [Google Scholar]
  16. Leder P., Bursztyn H. Initiation of protein synthesis,I. Effect of formylation of methionyl-tRNA on codon recognition. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1579–1585. doi: 10.1073/pnas.56.5.1579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Maki H., Horiuchi T., Sekiguchi M. Structure and expression of the dnaQ mutator and the RNase H genes of Escherichia coli: overlap of the promoter regions. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7137–7141. doi: 10.1073/pnas.80.23.7137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nomura T., Aiba H., Ishihama A. Transcriptional organization of the convergent overlapping dnaQ-rnh genes of Escherichia coli. J Biol Chem. 1985 Jun 10;260(11):7122–7125. [PubMed] [Google Scholar]
  19. Nomura T., Fujita N., Ishihama A. Promoter selectivity of E. coli RNA polymerase: analysis of the promoter system of convergently-transcribed dnaQ-rnh genes. Nucleic Acids Res. 1985 Nov 11;13(21):7647–7661. doi: 10.1093/nar/13.21.7647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nomura T., Ishihama A., Kajitani M., Takahashi T., Nakada N., Yoshinaga K. Promoter selectivity of Escherichia coli RNA polymerase. II: Altered promoter selection by mutant holoenzymes. Mol Gen Genet. 1984;193(1):8–16. doi: 10.1007/BF00327407. [DOI] [PubMed] [Google Scholar]
  21. Pedersen S., Skouv J., Kajitani M., Ishihama A. Transcriptional organization of the rpsA operon of Escherichia coli. Mol Gen Genet. 1984;196(1):135–140. doi: 10.1007/BF00334105. [DOI] [PubMed] [Google Scholar]
  22. Pongs O., Ulbrich N. Specific binding of formylated initiator-tRNA to Escherichia coli RNA polymerase. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3064–3067. doi: 10.1073/pnas.73.9.3064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Putney S. D., Meléndez D. L., Schimmel P. R. Cloning, partial sequencing, and in vitro transcription of the gene for alanine tRNA synthetase. J Biol Chem. 1981 Jan 10;256(1):205–211. [PubMed] [Google Scholar]
  24. Putney S. D., Royal N. J., Neuman de Vegvar H., Herlihy W. C., Biemann K., Schimmel P. Primary structure of a large aminoacyl-tRNA synthetase. Science. 1981 Sep 25;213(4515):1497–1501. doi: 10.1126/science.7025207. [DOI] [PubMed] [Google Scholar]
  25. Shih A. Y., Eisenstadt J., Lengyel P. On the relation between ribonucleic acid synthesis and peptide chain initiation in E. coli. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1599–1605. doi: 10.1073/pnas.56.5.1599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Spassky A., Busby S. J., Danchin A., Buc H. On the binding of tRNA to Escherichia coli RNA polymerase. Eur J Biochem. 1979 Aug 15;99(1):187–201. doi: 10.1111/j.1432-1033.1979.tb13245.x. [DOI] [PubMed] [Google Scholar]
  27. TISSIERES A., BOURGEOIS S., GROS F. Inhibition of RNA polymerase by RNA. J Mol Biol. 1963 Jul;7:100–103. doi: 10.1016/s0022-2836(63)80024-8. [DOI] [PubMed] [Google Scholar]
  28. Travers A. RNA polymerase specificity and the control of growth. Nature. 1976 Oct 21;263(5579):641–646. doi: 10.1038/263641a0. [DOI] [PubMed] [Google Scholar]
  29. Yamao F., Inokuchi H., Cheung A., Ozeki H., Söll D. Escherichia coli glutaminyl-tRNA synthetase. I. Isolation and DNA sequence of the glnS gene. J Biol Chem. 1982 Oct 10;257(19):11639–11643. [PubMed] [Google Scholar]
  30. Yoshimura M., Inokuchi H., Ozeki H. Identification of transfer RNA suppressors in Escherichia coli. IV. Amber suppressor Su+6 a double mutant of a new species of leucine tRNA. J Mol Biol. 1984 Aug 25;177(4):627–644. doi: 10.1016/0022-2836(84)90041-x. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES