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. 1999 Aug 1;27(15):3173–3182. doi: 10.1093/nar/27.15.3173

Transcriptional pause, arrest and termination sites for RNA polymerase II in mammalian N- and c-myc genes.

R G Keene 1, A Mueller 1, R Landick 1, L London 1
PMCID: PMC148545  PMID: 10454615

Abstract

Using either highly purified RNA polymerase II (pol II) elongation complexes assembled on oligo(dC)-tailed templates or promoter-initiated (extract-generated) pol II elongation complexes, the precise 3" ends of transcripts produced during transcription in vitro at several human c- and N- myc pause, arrest and termination sites were determined. Despite a low overall similarity between the entire c- and N- myc first exon sequences, many positions of pol II pausing, arrest or termination occurred within short regions of related sequence shared between the c- and N- myc templates. The c- and N- myc genes showed three general classes of sequence conservation near intrinsic pause, arrest or termination sites: (i) sites where arrest or termination occurred after the synthesis of runs of uridines (Us) preceding the transcript 3" end, (ii) sites downstream of potential RNA hairpins and (iii) sites after nucleotide addition following either a U or a C or following a combination of several pyrimidines near the transcript 3" end. The finding that regions of similarity occur near the sites of pol II pausing, arrest or termination suggests that the mechanism of c- and N- myc regulation at the level of transcript elongation may be similar and not divergent as previously proposed.

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Selected References

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

  1. Bengal E., Aloni Y. A block of transcription elongation by RNA polymerase II at synthetic sites in vitro. J Biol Chem. 1989 Jun 15;264(17):9791–9798. [PubMed] [Google Scholar]
  2. Bengal E., Flores O., Krauskopf A., Reinberg D., Aloni Y. Role of the mammalian transcription factors IIF, IIS, and IIX during elongation by RNA polymerase II. Mol Cell Biol. 1991 Mar;11(3):1195–1206. doi: 10.1128/mcb.11.3.1195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bentley D. L., Groudine M. A block to elongation is largely responsible for decreased transcription of c-myc in differentiated HL60 cells. Nature. 1986 Jun 12;321(6071):702–706. doi: 10.1038/321702a0. [DOI] [PubMed] [Google Scholar]
  4. Bentley D. L., Groudine M. Sequence requirements for premature termination of transcription in the human c-myc gene. Cell. 1988 Apr 22;53(2):245–256. doi: 10.1016/0092-8674(88)90386-8. [DOI] [PubMed] [Google Scholar]
  5. Bradsher J. N., Jackson K. W., Conaway R. C., Conaway J. W. RNA polymerase II transcription factor SIII. I. Identification, purification, and properties. J Biol Chem. 1993 Dec 5;268(34):25587–25593. [PubMed] [Google Scholar]
  6. Chamberlin M. J. New models for the mechanism of transcription elongation and its regulation. Harvey Lect. 1992 1993;88:1–21. [PubMed] [Google Scholar]
  7. Chan C. L., Landick R. Dissection of the his leader pause site by base substitution reveals a multipartite signal that includes a pause RNA hairpin. J Mol Biol. 1993 Sep 5;233(1):25–42. doi: 10.1006/jmbi.1993.1482. [DOI] [PubMed] [Google Scholar]
  8. Chan C. L., Landick R. The Salmonella typhimurium his operon leader region contains an RNA hairpin-dependent transcription pause site. Mechanistic implications of the effect on pausing of altered RNA hairpins. J Biol Chem. 1989 Dec 5;264(34):20796–20804. [PubMed] [Google Scholar]
  9. Dedrick R. L., Kane C. M., Chamberlin M. J. Purified RNA polymerase II recognizes specific termination sites during transcription in vitro. J Biol Chem. 1987 Jul 5;262(19):9098–9108. [PubMed] [Google Scholar]
  10. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Eick D., Bornkamm G. W. Transcriptional arrest within the first exon is a fast control mechanism in c-myc gene expression. Nucleic Acids Res. 1986 Nov 11;14(21):8331–8346. doi: 10.1093/nar/14.21.8331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Garrett K. P., Tan S., Bradsher J. N., Lane W. S., Conaway J. W., Conaway R. C. Molecular cloning of an essential subunit of RNA polymerase II elongation factor SIII. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5237–5241. doi: 10.1073/pnas.91.12.5237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Geiduschek E. P., Tocchini-Valentini G. P. Transcription by RNA polymerase III. Annu Rev Biochem. 1988;57:873–914. doi: 10.1146/annurev.bi.57.070188.004301. [DOI] [PubMed] [Google Scholar]
  14. Giardina C., Pérez-Riba M., Lis J. T. Promoter melting and TFIID complexes on Drosophila genes in vivo. Genes Dev. 1992 Nov;6(11):2190–2200. doi: 10.1101/gad.6.11.2190. [DOI] [PubMed] [Google Scholar]
  15. Gu W., Powell W., Mote J., Jr, Reines D. Nascent RNA cleavage by arrested RNA polymerase II does not require upstream translocation of the elongation complex on DNA. J Biol Chem. 1993 Dec 5;268(34):25604–25616. [PMC free article] [PubMed] [Google Scholar]
  16. Hawley D. K., Wiest D. K., Holtz M. S., Wang D. Transcriptional pausing, arrest, and readthrough at the adenovirus major late attenuation site. Cell Mol Biol Res. 1993;39(4):339–348. [PubMed] [Google Scholar]
  17. Izban M. G., Luse D. S. Factor-stimulated RNA polymerase II transcribes at physiological elongation rates on naked DNA but very poorly on chromatin templates. J Biol Chem. 1992 Jul 5;267(19):13647–13655. [PubMed] [Google Scholar]
  18. Izban M. G., Luse D. S. The RNA polymerase II ternary complex cleaves the nascent transcript in a 3'----5' direction in the presence of elongation factor SII. Genes Dev. 1992 Jul;6(7):1342–1356. doi: 10.1101/gad.6.7.1342. [DOI] [PubMed] [Google Scholar]
  19. Izban M. G., Luse D. S. The increment of SII-facilitated transcript cleavage varies dramatically between elongation competent and incompetent RNA polymerase II ternary complexes. J Biol Chem. 1993 Jun 15;268(17):12874–12885. [PubMed] [Google Scholar]
  20. Johnson T. L., Chamberlin M. J. Complexes of yeast RNA polymerase II and RNA are substrates for TFIIS-induced RNA cleavage. Cell. 1994 Apr 22;77(2):217–224. doi: 10.1016/0092-8674(94)90314-x. [DOI] [PubMed] [Google Scholar]
  21. Kash S. F., Kellems R. E. Control of transcription arrest in intron 1 of the murine adenosine deaminase gene. Mol Cell Biol. 1994 Sep;14(9):6198–6207. doi: 10.1128/mcb.14.9.6198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kerppola T. K., Kane C. M. Analysis of the signals for transcription termination by purified RNA polymerase II. Biochemistry. 1990 Jan 9;29(1):269–278. doi: 10.1021/bi00453a037. [DOI] [PubMed] [Google Scholar]
  23. Kerppola T. K., Kane C. M. Intrinsic sites of transcription termination and pausing in the c-myc gene. Mol Cell Biol. 1988 Oct;8(10):4389–4394. doi: 10.1128/mcb.8.10.4389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kerppola T. K., Kane C. M. RNA polymerase: regulation of transcript elongation and termination. FASEB J. 1991 Oct;5(13):2833–2842. doi: 10.1096/fasebj.5.13.1916107. [DOI] [PubMed] [Google Scholar]
  25. Kohl N. E., Legouy E., DePinho R. A., Nisen P. D., Smith R. K., Gee C. E., Alt F. W. Human N-myc is closely related in organization and nucleotide sequence to c-myc. Nature. 1986 Jan 2;319(6048):73–77. doi: 10.1038/319073a0. [DOI] [PubMed] [Google Scholar]
  26. Krumm A., Groudine M. Tumor suppression and transcription elongation: the dire consequences of changing partners. Science. 1995 Sep 8;269(5229):1400–1401. doi: 10.1126/science.7660121. [DOI] [PubMed] [Google Scholar]
  27. Krumm A., Meulia T., Brunvand M., Groudine M. The block to transcriptional elongation within the human c-myc gene is determined in the promoter-proximal region. Genes Dev. 1992 Nov;6(11):2201–2213. doi: 10.1101/gad.6.11.2201. [DOI] [PubMed] [Google Scholar]
  28. Krumm A., Meulia T., Groudine M. Common mechanisms for the control of eukaryotic transcriptional elongation. Bioessays. 1993 Oct;15(10):659–665. doi: 10.1002/bies.950151005. [DOI] [PubMed] [Google Scholar]
  29. Landick R., Roberts J. W. The shrewd grasp of RNA polymerase. Science. 1996 Jul 12;273(5272):202–203. doi: 10.1126/science.273.5272.202. [DOI] [PubMed] [Google Scholar]
  30. London L., Keene R. G., Landick R. Analysis of premature termination in c-myc during transcription by RNA polymerase II in a HeLa nuclear extract. Mol Cell Biol. 1991 Sep;11(9):4599–4615. doi: 10.1128/mcb.11.9.4599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Meulia T., Krumm A., Groudine M. Distinct properties of c-myc transcriptional elongation are revealed in Xenopus oocytes and mammalian cells and by template titration, 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), and promoter mutagenesis. Mol Cell Biol. 1993 Sep;13(9):5647–5658. doi: 10.1128/mcb.13.9.5647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Palangat M., Meier T. I., Keene R. G., Landick R. Transcriptional pausing at +62 of the HIV-1 nascent RNA modulates formation of the TAR RNA structure. Mol Cell. 1998 Jun;1(7):1033–1042. doi: 10.1016/s1097-2765(00)80103-3. [DOI] [PubMed] [Google Scholar]
  33. Resnekov O., Kessler M., Aloni Y. RNA secondary structure is an integral part of the in vitro mechanism of attenuation in simian virus 40. J Biol Chem. 1989 Jun 15;264(17):9953–9959. [PubMed] [Google Scholar]
  34. Samkurashvili I., Luse D. S. Translocation and transcriptional arrest during transcript elongation by RNA polymerase II. J Biol Chem. 1996 Sep 20;271(38):23495–23505. doi: 10.1074/jbc.271.38.23495. [DOI] [PubMed] [Google Scholar]
  35. Shilatifard A., Conaway J. W., Conaway R. C. Mechanism and regulation of transcriptional elongation and termination by RNA polymerase II. Curr Opin Genet Dev. 1997 Apr;7(2):199–204. doi: 10.1016/s0959-437x(97)80129-3. [DOI] [PubMed] [Google Scholar]
  36. Sluder A. E., Greenleaf A. L., Price D. H. Properties of a Drosophila RNA polymerase II elongation factor. J Biol Chem. 1989 May 25;264(15):8963–8969. [PubMed] [Google Scholar]
  37. Spencer C. A., Groudine M. Transcription elongation and eukaryotic gene regulation. Oncogene. 1990 Jun;5(6):777–785. [PubMed] [Google Scholar]
  38. Spencer C. A., Kilvert M. A. Transcription elongation in the human c-myc gene is governed by overall transcription initiation levels in Xenopus oocytes. Mol Cell Biol. 1993 Feb;13(2):1296–1305. doi: 10.1128/mcb.13.2.1296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Thompson N. E., Aronson D. B., Burgess R. R. Purification of eukaryotic RNA polymerase II by immunoaffinity chromatography. Elution of active enzyme with protein stabilizing agents from a polyol-responsive monoclonal antibody. J Biol Chem. 1990 Apr 25;265(12):7069–7077. [PubMed] [Google Scholar]
  40. Uptain S. M., Kane C. M., Chamberlin M. J. Basic mechanisms of transcript elongation and its regulation. Annu Rev Biochem. 1997;66:117–172. doi: 10.1146/annurev.biochem.66.1.117. [DOI] [PubMed] [Google Scholar]
  41. Wiest D. K., Wang D., Hawley D. K. Mechanistic studies of transcription arrest at the adenovirus major late attenuation site. Comparison of purified RNA polymerase II and washed elongation complexes. J Biol Chem. 1992 Apr 15;267(11):7733–7744. [PubMed] [Google Scholar]
  42. Wright S., Bishop J. M. DNA sequences that mediate attenuation of transcription from the mouse protooncogene myc. Proc Natl Acad Sci U S A. 1989 Jan;86(2):505–509. doi: 10.1073/pnas.86.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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]
  44. Xu L., Meng Y., Wallen R., DePinho R. A. Loss of transcriptional attenuation in N-myc is associated with progression towards a more malignant phenotype. Oncogene. 1995 Nov 2;11(9):1865–1872. [PubMed] [Google Scholar]

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