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. 1979 Oct;76(10):5110–5113. doi: 10.1073/pnas.76.10.5110

Termination of transcription by bacteriophage T3 RNA polymerase: homogeneous 3'-terminal oligonucleotide sequence of in vitro T3 RNA polymerase transcripts.

H K Majumder, U Maitra, M Rosenberg
PMCID: PMC413089  PMID: 388430

Abstract

RNA was synthesized in vitro from a T3 DNA template by T3 RNA polymerase and subsequently separated into seven discrete size classes (molecular weights ranging between 0.21 x 10(6) and 6.2 x 10(6)) by electrophoresis in polyacrylamide slab gels. RNase T1-generated 3'-terminal oligonucleotide fragments were then selectively isolated from either the unfractionated total RNA or the gel-purified specific transcripts by chromatography on columns of dihydroxyboryl-cellulose. Sequence analysis of these oligonucleotide products indicated that the unfractionated transcripts as well as all the individual major RNA species examined had a unique sequence, (Gp)UpUpUpUpUpGOH, at their 3' termini. The specificity of this sequence, as well as the total lack of any sequence heterogeneity at the ends of these transcripts, indicates a high degree of specificity of termination during transcription in this system.

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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., Sarkar P., Valenzuela D., Maitra U. Termination of transcription by Escherichia coli RNA polymerase: influence of secondary structure of RNA transcripts on rho-independent and rho-dependent termination. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1613–1617. doi: 10.1073/pnas.76.4.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beier H., Golomb M., Chamberlin M. Isolation of recombinants between T7 and T3 bacteriophages and their use in vitro transcriptional mapping. J Virol. 1977 Feb;21(2):753–765. doi: 10.1128/jvi.21.2.753-765.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bertrand K., Korn L., Lee F., Platt T., Squires C. L., Squires C., Yanofsky C. New features of the regulation of the tryptophan operon. Science. 1975 Jul 4;189(4196):22–26. doi: 10.1126/science.1094538. [DOI] [PubMed] [Google Scholar]
  5. Brownlee G. G., Sanger F. Chromatography of 32P-labelled oligonucleotides on thin layers of DEAE-cellulose. Eur J Biochem. 1969 Dec;11(2):395–399. doi: 10.1111/j.1432-1033.1969.tb00786.x. [DOI] [PubMed] [Google Scholar]
  6. Chakraborty P. R., Bandyopadhyay P., Huang H. H., Maitra U. Fidelity of in vitro transcription of T3 deoxyribonucleic acid by bacteriophage T3-induced ribonucleic acid polymerase and by Escherichia coli ribonucleic acid polymerase. J Biol Chem. 1974 Nov 10;249(21):6901–6909. [PubMed] [Google Scholar]
  7. Chakraborty P. R., Salvo R. A., Majumder H. K., Maitra U. Further characterization of bacteriophage T3-induced ribonucleic acid polymerase. Studies on the size of in vitro transcripts and interaction of T3 RNA polymerase with T3 DNA. J Biol Chem. 1977 Sep 25;252(18):6485–6493. [PubMed] [Google Scholar]
  8. Chakraborty P. R., Sarkar P., Huang H. H., Maitra U. Studies on T3-induced ribonucleic acid polymerase. 3. Purification and characterization of the T3-induced ribonucleic acid polymerase from bacteriophage T3-infected Escherichia coli cells. J Biol Chem. 1973 Oct 10;248(19):6637–6646. [PubMed] [Google Scholar]
  9. Dunn J. J., Bautz F. A., Bautz E. K. Different template specificities of phage T3 and T7 RNA polymerases. Nat New Biol. 1971 Mar 17;230(11):94–96. doi: 10.1038/newbio230094a0. [DOI] [PubMed] [Google Scholar]
  10. Dunn J. J., McAllister W. T., Bautz E. K. In vitro transcription of T3 DNA by Escherichia coli and T3 polymerases. Virology. 1972 Apr;48(1):112–125. doi: 10.1016/0042-6822(72)90119-5. [DOI] [PubMed] [Google Scholar]
  11. Franklin R. M. Purification and properties of the replicative intermediate of the RNA bacteriophage R17. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1504–1511. doi: 10.1073/pnas.55.6.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Golomb M., Chamberlin M. J. T7- and T3-specific RNA polymerases: characterization and mapping of the in vitro transcripts read from T3 DNA. J Virol. 1977 Feb;21(2):743–752. doi: 10.1128/jvi.21.2.743-752.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Küpper H., Sekiya T., Rosenberg M., Egan J., Landy A. A rho-dependent termination site in the gene coding for tyrosine tRNA su3 of Escherichia coli. Nature. 1978 Mar 30;272(5652):423–428. doi: 10.1038/272423a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lebowitz P., Weissman S. M., Radding C. M. Nucleotide sequence of a ribonucleic acid transcribed in vitro from lambda phage deoxyribonucleic acid. J Biol Chem. 1971 Aug 25;246(16):5120–5139. [PubMed] [Google Scholar]
  15. Lee F., Yanofsky C. Transcription termination at the trp operon attenuators of Escherichia coli and Salmonella typhimurium: RNA secondary structure and regulation of termination. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4365–4369. doi: 10.1073/pnas.74.10.4365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maitra U., Huang H. H. Initiation, release, and reinitiation of RNA chains by bacteriophage-T3-induced polymerase from T3 DNA templates (E. coli-guanosine triphosphate terminus-purified polymerase). Proc Natl Acad Sci U S A. 1972 Jan;69(1):55–59. doi: 10.1073/pnas.69.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Maitra U. Induction of a new RNA polymerase in Escherichia coli infected with bacteriophage T3. Biochem Biophys Res Commun. 1971 Apr 16;43(2):443–450. doi: 10.1016/0006-291x(71)90773-x. [DOI] [PubMed] [Google Scholar]
  18. Maitra U., Salvo R. A., Chakraborty P. R. Specificity of ribonucleic acid chain initiation by bacteriophage T3-induced ribonucleic acid polymerase. J Biol Chem. 1974 Sep 25;249(18):5835–5839. [PubMed] [Google Scholar]
  19. McAllister W. T., Küpper H., Bautz E. K. Kinetics of transcription by the bacteriophage-T3 RNA polymerase in vitro. Eur J Biochem. 1973 May 2;34(3):489–501. doi: 10.1111/j.1432-1033.1973.tb02785.x. [DOI] [PubMed] [Google Scholar]
  20. Neff N. F., Chamberlin M. J. Termination of transcription by Escherichia coli RNA polymerase in vitro is affected by ribonucleoside triphosphate base analogs. J Biol Chem. 1978 Apr 10;253(7):2455–2460. [PubMed] [Google Scholar]
  21. Pieczenik G., Barrell B. G., Gefter M. L. Bacteriophage phi 80-induced low molecular weight RNA. Arch Biochem Biophys. 1972 Sep;152(1):152–165. doi: 10.1016/0003-9861(72)90203-2. [DOI] [PubMed] [Google Scholar]
  22. Rosenberg M., Court D., Shimatake H., Brady C., Wulff D. L. The relationship between function and DNA sequence in an intercistronic regulatory region in phage lambda. Nature. 1978 Mar 30;272(5652):414–423. doi: 10.1038/272414a0. [DOI] [PubMed] [Google Scholar]
  23. Rosenberg M. Isolation and sequence determination of the 3'-terminal regions of isotopically labelled RNA molecules. Nucleic Acids Res. 1974 May;1(5):653–671. doi: 10.1093/nar/1.5.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rosenberg M., Weissman S., deCrombrugghe B. Termination of transcription in bacteriophage lambda. Heterogeneous, 3'-terminal oligo-adenylate additions and the effects of rho factor. J Biol Chem. 1975 Jun 25;250(12):4755–4764. [PubMed] [Google Scholar]
  25. Rosenberg M., de Chrombrugghe B., Musso R. Determination of nucleotide sequences beyond the sites of transcriptional termination. Proc Natl Acad Sci U S A. 1976 Mar;73(3):717–721. doi: 10.1073/pnas.73.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sanger F., Brownlee G. G., Barrell B. G. A two-dimensional fractionation procedure for radioactive nucleotides. J Mol Biol. 1965 Sep;13(2):373–398. doi: 10.1016/s0022-2836(65)80104-8. [DOI] [PubMed] [Google Scholar]
  27. Sogin M. L., Pace N. R. Nucleotide sequence of 5 S ribosomal RNA precursor from Bacillus subtilis. J Biol Chem. 1976 Jun 10;251(11):3480–3488. [PubMed] [Google Scholar]
  28. Weith H. L., Wiebers J. L., Gilham P. T. Synthesis of cellulose derivatives containing the dihydroxyboryl group and a study of their capacity to form specific complexes with sugars and nucleic acid components. Biochemistry. 1970 Oct 27;9(22):4396–4401. doi: 10.1021/bi00824a021. [DOI] [PubMed] [Google Scholar]

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