Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1997 Jan 2;16(1):163–172. doi: 10.1093/emboj/16.1.163

Identification of a transcript release activity acting on ternary transcription complexes containing murine RNA polymerase I.

S W Mason 1, E E Sander 1, I Grummt 1
PMCID: PMC1169623  PMID: 9009277

Abstract

Termination of mammalian ribosomal gene transcription by RNA polymerase I (Pol I) requires binding of the nucleolar factor TTF-I (transcription termination factor for Pol I) to specific rDNA terminator elements. We have used recombinant murine TTF-I in an immobilized tailed template assay to analyze individual steps of the termination reaction. We demonstrate that, besides the TTF-I-DNA complex which stops elongating Pol I, an additional activity is required to release both the nascent transcript and Pol I from the template. Moreover, transcript release, but not TTF-I-directed pausing, depends on upstream sequences directly flanking the terminator element. Together, complete termination of Pol I transcription requires TTF-I bound to the terminator DNA, a stretch of thymidine residues upstream of the TTF-I-mediated pause site and an activity which releases the RNA transcript and Pol I from the DNA template.

Full Text

The Full Text of this article is available as a PDF (357.0 KB).

Selected References

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

  1. Dieci G., Sentenac A. Facilitated recycling pathway for RNA polymerase III. Cell. 1996 Jan 26;84(2):245–252. doi: 10.1016/s0092-8674(00)80979-4. [DOI] [PubMed] [Google Scholar]
  2. Gottlieb E., Steitz J. A. Function of the mammalian La protein: evidence for its action in transcription termination by RNA polymerase III. EMBO J. 1989 Mar;8(3):851–861. doi: 10.1002/j.1460-2075.1989.tb03446.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Grummt I., Kuhn A., Bartsch I., Rosenbauer H. A transcription terminator located upstream of the mouse rDNA initiation site affects rRNA synthesis. Cell. 1986 Dec 26;47(6):901–911. doi: 10.1016/0092-8674(86)90805-6. [DOI] [PubMed] [Google Scholar]
  4. Grummt I., Rosenbauer H., Niedermeyer I., Maier U., Ohrlein A. A repeated 18 bp sequence motif in the mouse rDNA spacer mediates binding of a nuclear factor and transcription termination. Cell. 1986 Jun 20;45(6):837–846. doi: 10.1016/0092-8674(86)90558-1. [DOI] [PubMed] [Google Scholar]
  5. Hinkle D. C., Ring J., Chamberlin M. J. Studies of the binding of Escherichia coli RNA polymerase to DNA. 3. Tight binding of RNA polymerase holoenzyme to single-strand breaks in T7 DNA. J Mol Biol. 1972 Sep 28;70(2):197–207. doi: 10.1016/0022-2836(72)90533-5. [DOI] [PubMed] [Google Scholar]
  6. Jeong S. W., Lang W. H., Reeder R. H. The release element of the yeast polymerase I transcription terminator can function independently of Reb1p. Mol Cell Biol. 1995 Nov;15(11):5929–5936. doi: 10.1128/mcb.15.11.5929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kuhn A., Bartsch I., Grummt I. Specific interaction of the murine transcription termination factor TTF I with class-I RNA polymerases. Nature. 1990 Apr 5;344(6266):559–562. doi: 10.1038/344559a0. [DOI] [PubMed] [Google Scholar]
  8. Kuhn A., Grummt I. 3'-end formation of mouse pre-rRNA involves both transcription termination and a specific processing reaction. Genes Dev. 1989 Feb;3(2):224–231. doi: 10.1101/gad.3.2.224. [DOI] [PubMed] [Google Scholar]
  9. Kuhn A., Normann A., Bartsch I., Grummt I. The mouse ribosomal gene terminator consists of three functionally separable sequence elements. EMBO J. 1988 May;7(5):1497–1502. doi: 10.1002/j.1460-2075.1988.tb02968.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Labhart P., Reeder R. H. Characterization of three sites of RNA 3' end formation in the Xenopus ribosomal gene spacer. Cell. 1986 May 9;45(3):431–443. doi: 10.1016/0092-8674(86)90329-6. [DOI] [PubMed] [Google Scholar]
  11. Labhart P., Reeder R. H. Ribosomal precursor 3' end formation requires a conserved element upstream of the promoter. Cell. 1987 Jul 3;50(1):51–57. doi: 10.1016/0092-8674(87)90661-1. [DOI] [PubMed] [Google Scholar]
  12. Labhart P. The Xenopus 9 bp ribosomal terminator (T3 box) is a pause signal for the RNA polymerase I elongation complex. Nucleic Acids Res. 1995 Jun 25;23(12):2252–2258. doi: 10.1093/nar/23.12.2252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lang W. H., Morrow B. E., Ju Q., Warner J. R., Reeder R. H. A model for transcription termination by RNA polymerase I. Cell. 1994 Nov 4;79(3):527–534. doi: 10.1016/0092-8674(94)90261-5. [DOI] [PubMed] [Google Scholar]
  14. Lang W. H., Reeder R. H. The REB1 site is an essential component of a terminator for RNA polymerase I in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Jan;13(1):649–658. doi: 10.1128/mcb.13.1.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lang W. H., Reeder R. H. Transcription termination of RNA polymerase I due to a T-rich element interacting with Reb1p. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9781–9785. doi: 10.1073/pnas.92.21.9781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lee D. N., Feng G., Landick R. GreA-induced transcript cleavage is accompanied by reverse translocation to a different transcription complex conformation. J Biol Chem. 1994 Sep 2;269(35):22295–22303. [PubMed] [Google Scholar]
  17. Li J., Mason S. W., Greenblatt J. Elongation factor NusG interacts with termination factor rho to regulate termination and antitermination of transcription. Genes Dev. 1993 Jan;7(1):161–172. doi: 10.1101/gad.7.1.161. [DOI] [PubMed] [Google Scholar]
  18. Maraia R. J. Transcription termination factor La is also an initiation factor for RNA polymerase III. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3383–3387. doi: 10.1073/pnas.93.8.3383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nudler E., Goldfarb A., Kashlev M. Discontinuous mechanism of transcription elongation. Science. 1994 Aug 5;265(5173):793–796. doi: 10.1126/science.8047884. [DOI] [PubMed] [Google Scholar]
  20. Nudler E., Kashlev M., Nikiforov V., Goldfarb A. Coupling between transcription termination and RNA polymerase inchworming. Cell. 1995 May 5;81(3):351–357. doi: 10.1016/0092-8674(95)90388-7. [DOI] [PubMed] [Google Scholar]
  21. Orlova M., Newlands J., Das A., Goldfarb A., Borukhov S. Intrinsic transcript cleavage activity of RNA polymerase. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4596–4600. doi: 10.1073/pnas.92.10.4596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Reines D., Mote J., Jr Elongation factor SII-dependent transcription by RNA polymerase II through a sequence-specific DNA-binding protein. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1917–1921. doi: 10.1073/pnas.90.5.1917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Richardson J. P. Transcription termination. Crit Rev Biochem Mol Biol. 1993;28(1):1–30. doi: 10.3109/10409239309082571. [DOI] [PubMed] [Google Scholar]
  25. Riva M., Buhler J. M., Sentenac A., Fromageot P., Hawthorne D. C. Natural variation in yeast RNA polymerase A. Formation of a mosaic RNA polymerase A in a meiotic segregant from an interspecific hybrid. J Biol Chem. 1982 Apr 25;257(8):4570–4577. [PubMed] [Google Scholar]
  26. Sander E. E., Mason S. W., Munz C., Grummt I. The amino-terminal domain of the transcription termination factor TTF-I causes protein oligomerization and inhibition of DNA binding. Nucleic Acids Res. 1996 Oct 1;24(19):3677–3684. doi: 10.1093/nar/24.19.3677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schnapp A., Grummt I. Purification, assay, and properties of RNA polymerase I and class I-specific transcription factors in mouse. Methods Enzymol. 1996;273:233–248. doi: 10.1016/s0076-6879(96)73023-9. [DOI] [PubMed] [Google Scholar]
  28. Schnapp G., Graveley B. R., Grummt I. TFIIS binds to mouse RNA polymerase I and stimulates transcript elongation and hydrolytic cleavage of nascent rRNA. Mol Gen Genet. 1996 Sep 25;252(4):412–419. doi: 10.1007/BF02173006. [DOI] [PubMed] [Google Scholar]
  29. Smid A., Finsterer M., Grummt I. Limited proteolysis unmasks specific DNA-binding of the murine RNA polymerase I-specific transcription termination factor TTFI. J Mol Biol. 1992 Oct 5;227(3):635–647. doi: 10.1016/0022-2836(92)90213-4. [DOI] [PubMed] [Google Scholar]
  30. Wang D., Meier T. I., Chan C. L., Feng G., Lee D. N., Landick R. Discontinuous movements of DNA and RNA in RNA polymerase accompany formation of a paused transcription complex. Cell. 1995 May 5;81(3):341–350. doi: 10.1016/0092-8674(95)90387-9. [DOI] [PubMed] [Google Scholar]
  31. Xie Z., Price D. H. Purification of an RNA polymerase II transcript release factor from Drosophila. J Biol Chem. 1996 May 10;271(19):11043–11046. doi: 10.1074/jbc.271.19.11043. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES