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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Nov;81(21):6789–6793. doi: 10.1073/pnas.81.21.6789

Antitermination of transcription from an Escherichia coli ribosomal RNA promoter.

W E Holben, E A Morgan
PMCID: PMC392017  PMID: 6208558

Abstract

The Escherichia coli lac and ara promoters and rrnC ribosomal RNA promoter-leader region were fused to lacZYA. Transcription termination signals were introduced into the lac genes of these fusions by Tn9 and IS1 insertions. Measurement of lac enzymes from upstream and downstream of the insertions showed that termination signals resulting from these insertions are very efficient when transcription begins at lac or ara promoters but are very inefficient when transcription begins at the rrnC promoter-leader region. The rrnC promoter-leader region must, therefore, modify RNA polymerase to enable it to read through transcription termination signals.

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Selected References

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  1. Adhya S., Gottesman M. Control of transcription termination. Annu Rev Biochem. 1978;47:967–996. doi: 10.1146/annurev.bi.47.070178.004535. [DOI] [PubMed] [Google Scholar]
  2. Adhya S., Gottesman M., De Crombrugghe B. Release of polarity in Escherichia coli by gene N of phage lambda: termination and antitermination of transcription. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2534–2538. doi: 10.1073/pnas.71.6.2534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aksoy S., Squires C. L., Squires C. Translational coupling of the trpB and trpA genes in the Escherichia coli tryptophan operon. J Bacteriol. 1984 Feb;157(2):363–367. doi: 10.1128/jb.157.2.363-367.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Alton N. K., Vapnek D. Nucleotide sequence analysis of the chloramphenicol resistance transposon Tn9. Nature. 1979 Dec 20;282(5741):864–869. doi: 10.1038/282864a0. [DOI] [PubMed] [Google Scholar]
  5. Besemer J., Herpers M. Suppression of polarity of insertion mutations within the gal operon of E. coli. Mol Gen Genet. 1977 Mar 16;151(3):295–304. doi: 10.1007/BF00268793. [DOI] [PubMed] [Google Scholar]
  6. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  7. Brewster J. M., Morgan E. A. Tn9 and IS1 inserts in a ribosomal ribonucleic acid operon of Escherichia coli are incompletely polar. J Bacteriol. 1981 Dec;148(3):897–903. doi: 10.1128/jb.148.3.897-903.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brosius J., Dull T. J., Sleeter D. D., Noller H. F. Gene organization and primary structure of a ribosomal RNA operon from Escherichia coli. J Mol Biol. 1981 May 15;148(2):107–127. doi: 10.1016/0022-2836(81)90508-8. [DOI] [PubMed] [Google Scholar]
  9. Calva E., Burgess R. R. Characterization of a rho-dependent termination site within the cro gene of bacteriophage lambda. J Biol Chem. 1980 Nov 25;255(22):11017–11022. [PubMed] [Google Scholar]
  10. Casadaban M. J., Cohen S. N. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. doi: 10.1016/0022-2836(80)90283-1. [DOI] [PubMed] [Google Scholar]
  11. De Crombrugghe B., Adhya S., Gottesman M., Pastan I. Effect of Rho on transcription of bacterial operons. Nat New Biol. 1973 Feb 28;241(113):260–264. doi: 10.1038/newbio241260a0. [DOI] [PubMed] [Google Scholar]
  12. Duester G. L., Holmes W. M. The distal end of the ribosomal RNA operon rrnD of Escherichia coli contains a tRNA1thr gene, two 5s rRNA genes and a transcription terminator. Nucleic Acids Res. 1980 Sep 11;8(17):3793–3807. doi: 10.1093/nar/8.17.3793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Forbes D., Herskowitz I. Polarity suppression by the Q gene product of bacteriophage lambda. J Mol Biol. 1982 Oct 5;160(4):549–569. doi: 10.1016/0022-2836(82)90314-x. [DOI] [PubMed] [Google Scholar]
  14. Friedman D. I., Olson E. R. Evidence that a nucleotide sequence, "boxA," is involved in the action of the NusA protein. Cell. 1983 Aug;34(1):143–149. doi: 10.1016/0092-8674(83)90144-7. [DOI] [PubMed] [Google Scholar]
  15. Grayhack E. J., Roberts J. W. The phage lambda Q gene product: activity of a transcription antiterminator in vitro. Cell. 1982 Sep;30(2):637–648. doi: 10.1016/0092-8674(82)90260-4. [DOI] [PubMed] [Google Scholar]
  16. Ikemura T., Nomura M. Expression of spacer tRNA genes in ribosomal RNA transcription units carried by hybrid Col E1 plasmids in E. coli. Cell. 1977 Aug;11(4):779–793. doi: 10.1016/0092-8674(77)90291-4. [DOI] [PubMed] [Google Scholar]
  17. Imamoto F. Evidence for premature termination of transcription of the tryptophan operon in polarity mutants of Escherichia coli. Nature. 1970 Oct 17;228(5268):232–235. doi: 10.1038/228232a0. [DOI] [PubMed] [Google Scholar]
  18. Kahn M., Kolter R., Thomas C., Figurski D., Meyer R., Remaut E., Helinski D. R. Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2. Methods Enzymol. 1979;68:268–280. doi: 10.1016/0076-6879(79)68019-9. [DOI] [PubMed] [Google Scholar]
  19. Kingsbury D. T., Helinski D. R. DNA polymerase as a requirement for the maintenance of the bacterial plasmid colicinogenic factor E1. Biochem Biophys Res Commun. 1970 Dec 24;41(6):1538–1544. doi: 10.1016/0006-291x(70)90562-0. [DOI] [PubMed] [Google Scholar]
  20. Kingston R. E., Chamberlin M. J. Pausing and attenuation of in vitro transcription in the rrnB operon of E. coli. Cell. 1981 Dec;27(3 Pt 2):523–531. doi: 10.1016/0092-8674(81)90394-9. [DOI] [PubMed] [Google Scholar]
  21. Miller J. H., Calos M. P., Galas D., Hofer M., Büchel D. E., Müller-Hill B. Genetic analysis of transpositions in the lac region of Escherichia coli. J Mol Biol. 1980 Nov 25;144(1):1–18. doi: 10.1016/0022-2836(80)90212-0. [DOI] [PubMed] [Google Scholar]
  22. Morgan E. A., Ikemura T., Lindahl L., Fallon A. M., Nomura M. Some rRNA operons in E. coli have tRNA genes at their distal ends. Cell. 1978 Feb;13(2):335–344. doi: 10.1016/0092-8674(78)90202-7. [DOI] [PubMed] [Google Scholar]
  23. Morgan E. A. Insertions of Tn 10 into an E. coli ribosomal RNA operon are incompletely polar. Cell. 1980 Aug;21(1):257–265. doi: 10.1016/0092-8674(80)90133-6. [DOI] [PubMed] [Google Scholar]
  24. Morse D. E., Guertin M. Amber suA mutations which relieve polarity. J Mol Biol. 1972 Feb 14;63(3):605–608. doi: 10.1016/0022-2836(72)90453-6. [DOI] [PubMed] [Google Scholar]
  25. Olson E. R., Flamm E. L., Friedman D. I. Analysis of nutR: a region of phage lambda required for antitermination of transcription. Cell. 1982 Nov;31(1):61–70. doi: 10.1016/0092-8674(82)90405-6. [DOI] [PubMed] [Google Scholar]
  26. Proctor G. N., Rownd R. H. Rosanilins: indicator dyes for chloramphenicol-resistant enterobacteria containing chloramphenicol acetyltransferase. J Bacteriol. 1982 Jun;150(3):1375–1382. doi: 10.1128/jb.150.3.1375-1382.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ratner D. Evidence that mutations in the suA polarity suppressing gene directly affect termination factor rho. Nature. 1976 Jan 15;259(5539):151–153. doi: 10.1038/259151a0. [DOI] [PubMed] [Google Scholar]
  28. Reyes O., Gottesman M., Adhya S. Suppression of polarity of insertion mutations in the gal operon and N mutations in bacteriophage lambda. J Bacteriol. 1976 Jun;126(3):1108–1112. doi: 10.1128/jb.126.3.1108-1112.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Richardson J. P., Grimley C., Lowery C. Transcription termination factor rho activity is altered in Escherichia coli with suA gene mutations. Proc Natl Acad Sci U S A. 1975 May;72(5):1725–1728. doi: 10.1073/pnas.72.5.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Segawa T., Imamoto F. Diversity of regulation of genetic transcription. II. Specific relaxation of polarity in read-through transcription of the translocated trp operon in bacteriophage lambda trp. J Mol Biol. 1974 Aug 25;87(4):741–754. doi: 10.1016/0022-2836(74)90082-5. [DOI] [PubMed] [Google Scholar]
  31. Sekiya T., Mori M., Takahashi N., Nishimura S. Sequence of the distal tRNA1Asp gene and the transcription termination signal in the Escherichia coli ribosomal RNA operon rrnF(or G). Nucleic Acids Res. 1980 Sep 11;8(17):3809–3827. doi: 10.1093/nar/8.17.3809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Siehnel R. J., Morgan E. A. Efficient read-through of Tn9 and IS1 by RNA polymerase molecules that initiate at rRNA promoters. J Bacteriol. 1983 Feb;153(2):672–684. doi: 10.1128/jb.153.2.672-684.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tsuchiya T., Ottina K., Moriyama Y., Newman M. J., Wilson T. H. Solubilization and reconstitution of the melibiose carrier from a plasmid-carrying strain of Escherichia coli. J Biol Chem. 1982 May 10;257(9):5125–5128. [PubMed] [Google Scholar]
  34. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  35. Ward D. F., Gottesman M. E. Suppression of transcription termination by phage lambda. Science. 1982 May 28;216(4549):946–951. doi: 10.1126/science.6281888. [DOI] [PubMed] [Google Scholar]
  36. Yanofsky C., Platt T., Crawford I. P., Nichols B. P., Christie G. E., Horowitz H., VanCleemput M., Wu A. M. The complete nucleotide sequence of the tryptophan operon of Escherichia coli. Nucleic Acids Res. 1981 Dec 21;9(24):6647–6668. doi: 10.1093/nar/9.24.6647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Young R. A. Transcription termination in the Escherichia coli ribosomal RNA operon rrnC. J Biol Chem. 1979 Dec 25;254(24):12725–12731. [PubMed] [Google Scholar]
  38. Zipser D., Zabell S., Rothman J., Grodzicker T., Wenk M. Fine structure of the gradient of polarity in the z gene of the lac operon of Escherichia coli. J Mol Biol. 1970 Apr 14;49(1):251–254. doi: 10.1016/0022-2836(70)90392-x. [DOI] [PubMed] [Google Scholar]

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