Skip to main content
NCBI home page
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 Dec;81(23):7373–7377. doi: 10.1073/pnas.81.23.7373

Identification and characterization of a new transcriptional termination factor from Escherichia coli.

J F Briat, M J Chamberlin
PMCID: PMC392148  PMID: 6095288

Abstract

We have identified and partially purified an activity from Escherichia coli that enhances transcription termination at the bacteriophage T7 early terminator when cloned on the plasmid pAR1707. The factor also causes the transcript to be terminated at a site several nucleotides earlier than in its absence. The resulting 3' OH ends of the transcripts are identical to those found in vivo by S1 nuclease mapping. From this we conclude that the factor we have identified is probably responsible for determination of the 3' OH ends of T7 RNA transcripts in vivo. This factor does not act by processing a preformed RNA transcript, nor is it replaced by rho protein or nusA or nusB proteins. Therefore, it appears to be a new transcription termination factor, and we have designated it "tau factor." Elucidation of its role in transcription in E. coli will depend on its purification to homogeneity and further studies of its properties.

Full text

PDF
7373

Images in this article

7375Image
on p.7375
7376Image
on p.7376
7377Image
on p.7377

Selected References

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

  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. 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]
  3. Burgess R. R., Jendrisak J. J. A procedure for the rapid, large-scall purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precipitation and DNA-cellulose chromatography. Biochemistry. 1975 Oct 21;14(21):4634–4638. doi: 10.1021/bi00692a011. [DOI] [PubMed] [Google Scholar]
  4. Chamberlin M. J., Nierman W. C., Wiggs J., Neff N. A quantitative assay for bacterial RNA polymerases. J Biol Chem. 1979 Oct 25;254(20):10061–10069. [PubMed] [Google Scholar]
  5. Davison B. L., Leighton T., Rabinowitz J. C. Purification of Bacillus subtilis RNA polymerase with heparin-agarose. In vitro transcription of phi 29 DNA. J Biol Chem. 1979 Sep 25;254(18):9220–9226. [PubMed] [Google Scholar]
  6. Dunn J. J., Studier F. W. The transcription termination site at the end of the early region of bacteriophage T7 DNA. Nucleic Acids Res. 1980 May 24;8(10):2119–2132. doi: 10.1093/nar/8.10.2119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fuller R. S., Kaguni J. M., Kornberg A. Enzymatic replication of the origin of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7370–7374. doi: 10.1073/pnas.78.12.7370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gilman M. Z., Chamberlin M. J. Developmental and genetic regulation of Bacillus subtilis genes transcribed by sigma 28-RNA polymerase. Cell. 1983 Nov;35(1):285–293. doi: 10.1016/0092-8674(83)90231-3. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  11. Howard B. H., de Crombrugghe B., Rosenberg M. Transcription in vitro of bacteriophage lambda 4S RNA: studies on termination and rho protein. Nucleic Acids Res. 1977 Apr;4(4):827–842. doi: 10.1093/nar/4.4.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kassavetis G. A., Chamberlin M. J. Pausing and termination of transcription within the early region of bacteriophage T7 DNA in vitro. J Biol Chem. 1981 Mar 25;256(6):2777–2786. [PubMed] [Google Scholar]
  13. Kiefer M., Neff N., Chamberlin M. J. Transcriptional termination at the end of the early region of bacteriophages T3 and T7 is not affected by polarity suppressors. J Virol. 1977 May;22(2):548–552. doi: 10.1128/jvi.22.2.548-552.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Korn L. J., Yanofsky C. Polarity suppressors defective in transcription termination at the attenuator of the tryptophan operon of Escherichia coli have altered rho factor. J Mol Biol. 1976 Sep 15;106(2):231–241. doi: 10.1016/0022-2836(76)90082-6. [DOI] [PubMed] [Google Scholar]
  16. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  17. McAllister W. T., Morris C., Rosenberg A. H., Studier F. W. Utilization of bacteriophage T7 late promoters in recombinant plasmids during infection. J Mol Biol. 1981 Dec 15;153(3):527–544. doi: 10.1016/0022-2836(81)90406-x. [DOI] [PubMed] [Google Scholar]
  18. Peters G. G., Hayward R. S. Dinucleotide sequences in the regions of T7 DNA coding for termination of early transcription. Eur J Biochem. 1974 Oct 1;48(1):199–208. doi: 10.1111/j.1432-1033.1974.tb03757.x. [DOI] [PubMed] [Google Scholar]
  19. Platt T. Termination of transcription and its regulation in the tryptophan operon of E. coli. Cell. 1981 Apr;24(1):10–23. doi: 10.1016/0092-8674(81)90496-7. [DOI] [PubMed] [Google Scholar]
  20. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  21. Schmidt M. C., Chamberlin M. J. Amplification and isolation of Escherichia coli nusA protein and studies of its effects on in vitro RNA chain elongation. Biochemistry. 1984 Jan 17;23(2):197–203. doi: 10.1021/bi00297a004. [DOI] [PubMed] [Google Scholar]
  22. Smith M., Leung D. W., Gillam S., Astell C. R., Montgomery D. L., Hall B. D. Sequence of the gene for iso-1-cytochrome c in Saccharomyces cerevisiae. Cell. 1979 Apr;16(4):753–761. doi: 10.1016/0092-8674(79)90091-6. [DOI] [PubMed] [Google Scholar]
  23. Studier F. W. Bacteriophage T7. Science. 1972 Apr 28;176(4033):367–376. doi: 10.1126/science.176.4033.367. [DOI] [PubMed] [Google Scholar]
  24. Symons R. H. The rapid, simple and improved preparation of high specific activity alpha-[32P]dATP and alpha-[32P]ATP. Nucleic Acids Res. 1977 Dec;4(12):4347–4355. doi: 10.1093/nar/4.12.4347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. von Hippel P. H., Bear D. G., Morgan W. D., McSwiggen J. A. Protein-nucleic acid interactions in transcription: a molecular analysis. Annu Rev Biochem. 1984;53:389–446. doi: 10.1146/annurev.bi.53.070184.002133. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES