Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1991 Apr 25;19(8):1845–1852. doi: 10.1093/nar/19.8.1845

Transcriptional termination sequence at the end of the Escherichia coli ribosomal RNA G operon: complex terminators and antitermination.

B Albrechtsen 1, B M Ross 1, C Squires 1, C L Squires 1
PMCID: PMC328114  PMID: 1709493

Abstract

We have examined the termination region sequence of the rrnG operon and have observed its properties in vivo using a fusion plasmid test system. Transcription of rrnG terminator fragments was also studied in vitro. We found that termination of rrnG transcription is a complex process controlled by a tandem Rho-independent and Rho-dependent terminator arrangement which we designate rrnG-tt'. Together, these two elements were 98% efficient at terminating transcription initiated at the rrnG-P2 promoter. When the two elements were separated, however, we found that the Rho-independent structure was only 59% efficient while the Rho-dependent fragment alone could account for total transcriptional termination of the tandem arrangement. The rrnG termination region was resistant to rrn antitermination and, therefore, possesses some means of stopping antiterminated transcription. The distal rrnG sequence contains several additional noteworthy features; the rrnGt' fragment contains a REP (repetitive extragenic palindromic) sequence and homology with a small unidentified reading frame following rrnE. This sequence is followed by witA, which is homologous to a citrate transport gene, citB. Finally, our sequence, obtained from plasmid pLC23-30, contains a Tn1000 insertion that is absent from the E. coli chromosome. This insertion lies 975 bp beyond the 5S gene and is not involved in the termination events examined in this study.

Full text

PDF
1845

Images in this article

Selected References

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

  1. Aksoy S., Squires C. L., Squires C. Evidence for antitermination in Escherichia coli RRNA transcription. J Bacteriol. 1984 Jul;159(1):260–264. doi: 10.1128/jb.159.1.260-264.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albrechtsen B., Squires C. L., Li S., Squires C. Antitermination of characterized transcriptional terminators by the Escherichia coli rrnG leader region. J Mol Biol. 1990 May 5;213(1):123–134. doi: 10.1016/S0022-2836(05)80125-1. [DOI] [PubMed] [Google Scholar]
  3. Barik S., Ghosh B., Whalen W., Lazinski D., Das A. An antitermination protein engages the elongating transcription apparatus at a promoter-proximal recognition site. Cell. 1987 Sep 11;50(6):885–899. doi: 10.1016/0092-8674(87)90515-0. [DOI] [PubMed] [Google Scholar]
  4. Berg K. L., Squires C. L., Squires C. In vivo translation of a region within the rrnB 16S rRNA gene of Escherichia coli. J Bacteriol. 1987 Apr;169(4):1691–1701. doi: 10.1128/jb.169.4.1691-1701.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berg K. L., Squires C., Squires C. L. Ribosomal RNA operon anti-termination. Function of leader and spacer region box B-box A sequences and their conservation in diverse micro-organisms. J Mol Biol. 1989 Oct 5;209(3):345–358. doi: 10.1016/0022-2836(89)90002-8. [DOI] [PubMed] [Google Scholar]
  6. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Chen C. Y., Richardson J. P. Sequence elements essential for rho-dependent transcription termination at lambda tR1. J Biol Chem. 1987 Aug 15;262(23):11292–11299. [PubMed] [Google Scholar]
  9. Clarke L., Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. doi: 10.1016/0092-8674(76)90055-6. [DOI] [PubMed] [Google Scholar]
  10. Das A., Court D., Adhya S. Isolation and characterization of conditional lethal mutants of Escherichia coli defective in transcription termination factor rho. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1959–1963. doi: 10.1073/pnas.73.6.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Ellwood M., Nomura M. Chromosomal locations of the genes for rRNA in Escherichia coli K-12. J Bacteriol. 1982 Feb;149(2):458–468. doi: 10.1128/jb.149.2.458-468.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Friedman D. I., Imperiale M. J., Adhya S. L. RNA 3' end formation in the control of gene expression. Annu Rev Genet. 1987;21:453–488. doi: 10.1146/annurev.ge.21.120187.002321. [DOI] [PubMed] [Google Scholar]
  14. Goliger J. A., Yang X. J., Guo H. C., Roberts J. W. Early transcribed sequences affect termination efficiency of Escherichia coli RNA polymerase. J Mol Biol. 1989 Jan 20;205(2):331–341. doi: 10.1016/0022-2836(89)90344-6. [DOI] [PubMed] [Google Scholar]
  15. Guterman S. K., Howitt C. L. Rifampicin supersensitivity of rho strains of E. coli, and suppression by sur mutation. Mol Gen Genet. 1979 Jan 16;169(1):27–34. doi: 10.1007/BF00267541. [DOI] [PubMed] [Google Scholar]
  16. Higgins C. F., McLaren R. S., Newbury S. F. Repetitive extragenic palindromic sequences, mRNA stability and gene expression: evolution by gene conversion? A review. Gene. 1988 Dec 10;72(1-2):3–14. doi: 10.1016/0378-1119(88)90122-9. [DOI] [PubMed] [Google Scholar]
  17. Horwitz R. J., Li J., Greenblatt J. An elongation control particle containing the N gene transcriptional antitermination protein of bacteriophage lambda. Cell. 1987 Nov 20;51(4):631–641. doi: 10.1016/0092-8674(87)90132-2. [DOI] [PubMed] [Google Scholar]
  18. Klotsky R. A., Schwartz I. Measurement of cat expression from growth-rate-regulated promoters employing beta-lactamase activity as an indicator of plasmid copy number. Gene. 1987;55(1):141–146. doi: 10.1016/0378-1119(87)90257-5. [DOI] [PubMed] [Google Scholar]
  19. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  20. Li S. C., Squires C. L., Squires C. Antitermination of E. coli rRNA transcription is caused by a control region segment containing lambda nut-like sequences. Cell. 1984 Oct;38(3):851–860. doi: 10.1016/0092-8674(84)90280-0. [DOI] [PubMed] [Google Scholar]
  21. Liebke H., Hatfull G. The sequence of the distal end of the E. coli ribosomal RNA rrnE operon indicates conserved features are shared by rrn operons. Nucleic Acids Res. 1985 Aug 12;13(15):5515–5525. doi: 10.1093/nar/13.15.5515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McSwiggen J. A., Bear D. G., von Hippel P. H. Interactions of Escherichia coli transcription termination factor rho with RNA. I. Binding stoichiometries and free energies. J Mol Biol. 1988 Feb 20;199(4):609–622. doi: 10.1016/0022-2836(88)90305-1. [DOI] [PubMed] [Google Scholar]
  23. Messing J., Gronenborn B., Müller-Hill B., Hans Hopschneider P. Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3642–3646. doi: 10.1073/pnas.74.9.3642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Morgan W. D., Bear D. G., Litchman B. L., von Hippel P. H. RNA sequence and secondary structure requirements for rho-dependent transcription termination. Nucleic Acids Res. 1985 May 24;13(10):3739–3754. doi: 10.1093/nar/13.10.3739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mott J. E., Galloway J. L., Platt T. Maturation of Escherichia coli tryptophan operon mRNA: evidence for 3' exonucleolytic processing after rho-dependent termination. EMBO J. 1985 Jul;4(7):1887–1891. doi: 10.1002/j.1460-2075.1985.tb03865.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Newbury S. F., Smith N. H., Robinson E. C., Hiles I. D., Higgins C. F. Stabilization of translationally active mRNA by prokaryotic REP sequences. Cell. 1987 Jan 30;48(2):297–310. doi: 10.1016/0092-8674(87)90433-8. [DOI] [PubMed] [Google Scholar]
  27. Rosenthal E. R., Calvo J. M. Transcription termination sites at the distal end of the leu operon of Salmonella typhimurium. J Mol Biol. 1987 Apr 5;194(3):443–452. doi: 10.1016/0022-2836(87)90673-5. [DOI] [PubMed] [Google Scholar]
  28. Saadi S., Maas W. K., Hill D. F., Bergquist P. L. Nucleotide sequence analysis of RepFIC, a basic replicon present in IncFI plasmids P307 and F, and its relation to the RepA replicon of IncFII plasmids. J Bacteriol. 1987 May;169(5):1836–1846. doi: 10.1128/jb.169.5.1836-1846.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sameshima J. H., Wek R. C., Hatfield G. W. Overlapping transcription and termination of the convergent ilvA and ilvY genes of Escherichia coli. J Biol Chem. 1989 Jan 15;264(2):1224–1231. [PubMed] [Google Scholar]
  30. Sasatsu M., Misra T. K., Chu L., Laddaga R., Silver S. Cloning and DNA sequence of a plasmid-determined citrate utilization system in Escherichia coli. J Bacteriol. 1985 Dec;164(3):983–993. doi: 10.1128/jb.164.3.983-993.1985. [DOI] [PMC free article] [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. Seol W., Shatkin A. J. A new gene located between pss and rrnG on the Escherichia coli chromosome. J Bacteriol. 1990 Sep;172(9):4745–4745. doi: 10.1128/jb.172.9.4745.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Seol W., Shatkin A. J. Sequence of the distal end of E. coli ribosomal RNA rrnG operon. Nucleic Acids Res. 1990 May 25;18(10):3056–3056. doi: 10.1093/nar/18.10.3056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shen W. F., Squires C., Squires C. L. Nucleotide sequence of the rrnG ribosomal RNA promoter region of Escherichia coli. Nucleic Acids Res. 1982 May 25;10(10):3303–3313. doi: 10.1093/nar/10.10.3303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Shyamala V., Schneider E., Ames G. F. Tandem chromosomal duplications: role of REP sequences in the recombination event at the join-point. EMBO J. 1990 Mar;9(3):939–946. doi: 10.1002/j.1460-2075.1990.tb08192.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Squires C., Krainer A., Barry G., Shen W. F., Squires C. L. Nucleotide sequence at the end of the gene for the RNA polymerase beta' subunit (rpoC). Nucleic Acids Res. 1981 Dec 21;9(24):6827–6840. doi: 10.1093/nar/9.24.6827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Szeberényi J., Apirion D. Initiation, processing and termination of ribosomal RNA from a hybrid 5 S ribosomal RNA gene in a plasmid. J Mol Biol. 1983 Aug 15;168(3):525–557. doi: 10.1016/s0022-2836(83)80300-3. [DOI] [PubMed] [Google Scholar]
  38. Telesnitsky A. P., Chamberlin M. J. Sequences linked to prokaryotic promoters can affect the efficiency of downstream termination sites. J Mol Biol. 1989 Jan 20;205(2):315–330. doi: 10.1016/0022-2836(89)90343-4. [DOI] [PubMed] [Google Scholar]
  39. Uhlin B. E., Nordström K. R plasmid gene dosage effects in Escherichia coli K-12: copy mutants of the R plasmic R1drd-19. Plasmid. 1977 Nov;1(1):1–7. doi: 10.1016/0147-619x(77)90003-8. [DOI] [PubMed] [Google Scholar]
  40. 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]

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

RESOURCES