Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1992 Mar;11(3):1085–1093. doi: 10.1002/j.1460-2075.1992.tb05147.x

Distinct modes of transcription read through or terminate at the c-myc attenuator.

S Roberts 1, D L Bentley 1
PMCID: PMC556549  PMID: 1372247

Abstract

Premature termination of transcription by RNA polymerase II (pol II) occurs in the 5' region of many viral and cellular genes. Modulation of this process, or attenuation, is an important means of transcriptional control, but its mechanism is unknown. Using injected Xenopus oocytes, the efficiency of the mouse c-myc attenuator was tested when it was placed at various distances from the transcription initiation site. The attenuator functioned with each of six different pol II promoters tested; however, termination efficiency declined markedly when it was placed more than approximately 400 bases from the start site. This decline in attenuator function with distance from the start site coincided with increased sensitivity to the pol II inhibitor 5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole (DRB). Thus transcription complexes situated further from the promoter appear to have a lower ability to recognize the attenuator and a greater sensitivity to DRB. Furthermore, polymerases which have read through one attenuation site have a reduced ability to terminate at a second site. The results imply that a discrete subset of elongation complexes is capable of premature termination, and that this subset exists only within the first few hundred bases of the transcription unit. Regulation of termination efficiency may be effected by changing the balance between the two modes of transcription committed either to read through or to terminate prematurely.

Full text

PDF
1085

Images in this article

1086Image
on p.1086
1087Image
on p.1087
1088Image
on p.1088
1089Image
on p.1089
1089Image
on p.1089
1090Image
on p.1090
1090Image
on p.1090

Selected References

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

  1. 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]
  2. Beaumont C., Porcher C., Picat C., Nordmann Y., Grandchamp B. The mouse porphobilinogen deaminase gene. Structural organization, sequence, and transcriptional analysis. J Biol Chem. 1989 Sep 5;264(25):14829–14834. [PubMed] [Google Scholar]
  3. Bender T. P., Thompson C. B., Kuehl W. M. Differential expression of c-myb mRNA in murine B lymphomas by a block to transcription elongation. Science. 1987 Sep 18;237(4821):1473–1476. doi: 10.1126/science.3498214. [DOI] [PubMed] [Google Scholar]
  4. Bentley D. L., Brown W. L., Groudine M. Accurate, TATA box-dependent polymerase III transcription from promoters of the c-myc gene in injected Xenopus oocytes. Genes Dev. 1989 Aug;3(8):1179–1189. doi: 10.1101/gad.3.8.1179. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Bentley D. L., Groudine M. Novel promoter upstream of the human c-myc gene and regulation of c-myc expression in B-cell lymphomas. Mol Cell Biol. 1986 Oct;6(10):3481–3489. doi: 10.1128/mcb.6.10.3481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Carbon P., Murgo S., Ebel J. P., Krol A., Tebb G., Mattaj L. W. A common octamer motif binding protein is involved in the transcription of U6 snRNA by RNA polymerase III and U2 snRNA by RNA polymerase II. Cell. 1987 Oct 9;51(1):71–79. doi: 10.1016/0092-8674(87)90011-0. [DOI] [PubMed] [Google Scholar]
  9. Chen Z., Harless M. L., Wright D. A., Kellems R. E. Identification and characterization of transcriptional arrest sites in exon 1 of the human adenosine deaminase gene. Mol Cell Biol. 1990 Sep;10(9):4555–4564. doi: 10.1128/mcb.10.9.4555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chinsky J. M., Maa M. C., Ramamurthy V., Kellems R. E. Adenosine deaminase gene expression. Tissue-dependent regulation of transcriptional elongation. J Biol Chem. 1989 Aug 25;264(24):14561–14565. [PubMed] [Google Scholar]
  11. Chodosh L. A., Fire A., Samuels M., Sharp P. A. 5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole inhibits transcription elongation by RNA polymerase II in vitro. J Biol Chem. 1989 Feb 5;264(4):2250–2257. [PubMed] [Google Scholar]
  12. Evans R., Weber J., Ziff E., Darnell J. E. Premature termination during adenovirus transcription. Nature. 1979 Mar 22;278(5702):367–370. doi: 10.1038/278367a0. [DOI] [PubMed] [Google Scholar]
  13. Fort P., Rech J., Vie A., Piechaczyk M., Bonnieu A., Jeanteur P., Blanchard J. M. Regulation of c-fos gene expression in hamster fibroblasts: initiation and elongation of transcription and mRNA degradation. Nucleic Acids Res. 1987 Jul 24;15(14):5657–5667. doi: 10.1093/nar/15.14.5657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fraser N. W., Sehgal P. B., Darnell J. E. DRB-induced premature termination of late adenovirus transcription. Nature. 1978 Apr 13;272(5654):590–593. doi: 10.1038/272590a0. [DOI] [PubMed] [Google Scholar]
  15. Fraser N. W., Sehgal P. B., Darnell J. E., Jr Multiple discrete sites for premature RNA chain termination late in adenovirus-2 infection: enhancement by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole. Proc Natl Acad Sci U S A. 1979 Jun;76(6):2571–2575. doi: 10.1073/pnas.76.6.2571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Grayhack E. J., Yang X. J., Lau L. F., Roberts J. W. Phage lambda gene Q antiterminator recognizes RNA polymerase near the promoter and accelerates it through a pause site. Cell. 1985 Aug;42(1):259–269. doi: 10.1016/s0092-8674(85)80121-5. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Haley J. D., Waterfield M. D. Contributory effects of de novo transcription and premature transcript termination in the regulation of human epidermal growth factor receptor proto-oncogene RNA synthesis. J Biol Chem. 1991 Jan 25;266(3):1746–1753. [PubMed] [Google Scholar]
  19. Hay N., Skolnik-David H., Aloni Y. Attenuation in the control of SV40 gene expression. Cell. 1982 May;29(1):183–193. doi: 10.1016/0092-8674(82)90102-7. [DOI] [PubMed] [Google Scholar]
  20. Henikoff S., Eghtedarzadeh M. K. Conserved arrangement of nested genes at the Drosophila Gart locus. Genetics. 1987 Dec;117(4):711–725. doi: 10.1093/genetics/117.4.711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hernandez N. Formation of the 3' end of U1 snRNA is directed by a conserved sequence located downstream of the coding region. EMBO J. 1985 Jul;4(7):1827–1837. doi: 10.1002/j.1460-2075.1985.tb03857.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Horikoshi M., Sekimizu K., Natori S. Analysis of the stimulatory factor of RNA polymerase II in the initiation and elongation complex. J Biol Chem. 1984 Jan 10;259(1):608–611. [PubMed] [Google Scholar]
  23. 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]
  24. Kao S. Y., Calman A. F., Luciw P. A., Peterlin B. M. Anti-termination of transcription within the long terminal repeat of HIV-1 by tat gene product. Nature. 1987 Dec 3;330(6147):489–493. doi: 10.1038/330489a0. [DOI] [PubMed] [Google Scholar]
  25. Krystal G., Birrer M., Way J., Nau M., Sausville E., Thompson C., Minna J., Battey J. Multiple mechanisms for transcriptional regulation of the myc gene family in small-cell lung cancer. Mol Cell Biol. 1988 Aug;8(8):3373–3381. doi: 10.1128/mcb.8.8.3373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Laspia M. F., Rice A. P., Mathews M. B. HIV-1 Tat protein increases transcriptional initiation and stabilizes elongation. Cell. 1989 Oct 20;59(2):283–292. doi: 10.1016/0092-8674(89)90290-0. [DOI] [PubMed] [Google Scholar]
  27. Logan J., Falck-Pedersen E., Darnell J. E., Jr, Shenk T. A poly(A) addition site and a downstream termination region are required for efficient cessation of transcription by RNA polymerase II in the mouse beta maj-globin gene. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8306–8310. doi: 10.1073/pnas.84.23.8306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lois R., Freeman L., Villeponteau B., Martinson H. G. Active beta-globin gene transcription occurs in methylated, DNase I-resistant chromatin of nonerythroid chicken cells. Mol Cell Biol. 1990 Jan;10(1):16–27. doi: 10.1128/mcb.10.1.16. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lund E., Dahlberg J. E. In vitro synthesis of vertebrate U1 snRNA. EMBO J. 1989 Jan;8(1):287–292. doi: 10.1002/j.1460-2075.1989.tb03375.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mechti N., Piechaczyk M., Blanchard J. M., Jeanteur P., Lebleu B. Sequence requirements for premature transcription arrest within the first intron of the mouse c-fos gene. Mol Cell Biol. 1991 May;11(5):2832–2841. doi: 10.1128/mcb.11.5.2832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nakamaye K. L., Eckstein F. Inhibition of restriction endonuclease Nci I cleavage by phosphorothioate groups and its application to oligonucleotide-directed mutagenesis. Nucleic Acids Res. 1986 Dec 22;14(24):9679–9698. doi: 10.1093/nar/14.24.9679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Nepveu A., Levine R. A., Campisi J., Greenberg M. E., Ziff E. B., Marcu K. B. Alternative modes of c-myc regulation in growth factor-stimulated and differentiating cells. Oncogene. 1987;1(3):243–250. [PubMed] [Google Scholar]
  33. Nepveu A., Marcu K. B. Intragenic pausing and anti-sense transcription within the murine c-myc locus. EMBO J. 1986 Nov;5(11):2859–2865. doi: 10.1002/j.1460-2075.1986.tb04580.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nepveu A., Marcu K. B., Skoultchi A. I., Lachman H. M. Contributions of transcriptional and post-transcriptional mechanisms to the regulation of c-myc expression in mouse erythroleukemia cells. Genes Dev. 1987 Nov;1(9):938–945. doi: 10.1101/gad.1.9.938. [DOI] [PubMed] [Google Scholar]
  35. Payne J. M., Laybourn P. J., Dahmus M. E. The transition of RNA polymerase II from initiation to elongation is associated with phosphorylation of the carboxyl-terminal domain of subunit IIa. J Biol Chem. 1989 Nov 25;264(33):19621–19629. [PubMed] [Google Scholar]
  36. Reinberg D., Roeder R. G. Factors involved in specific transcription by mammalian RNA polymerase II. Transcription factor IIS stimulates elongation of RNA chains. J Biol Chem. 1987 Mar 5;262(7):3331–3337. [PubMed] [Google Scholar]
  37. Rodaway A. R., Teahan C. G., Casimir C. M., Segal A. W., Bentley D. L. Characterization of the 47-kilodalton autosomal chronic granulomatous disease protein: tissue-specific expression and transcriptional control by retinoic acid. Mol Cell Biol. 1990 Oct;10(10):5388–5396. doi: 10.1128/mcb.10.10.5388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Skarnes W. C., Tessier D. C., Acheson N. H. RNA polymerases stall and/or prematurely terminate nearby both early and late promoters on polyomavirus DNA. J Mol Biol. 1988 Sep 5;203(1):153–171. doi: 10.1016/0022-2836(88)90099-x. [DOI] [PubMed] [Google Scholar]
  39. Spencer C. A., Groudine M. Transcription elongation and eukaryotic gene regulation. Oncogene. 1990 Jun;5(6):777–785. [PubMed] [Google Scholar]
  40. Spencer C. A., LeStrange R. C., Novak U., Hayward W. S., Groudine M. The block to transcription elongation is promoter dependent in normal and Burkitt's lymphoma c-myc alleles. Genes Dev. 1990 Jan;4(1):75–88. doi: 10.1101/gad.4.1.75. [DOI] [PubMed] [Google Scholar]
  41. Stevens A., Maupin M. K. 5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole inhibits a HeLa protein kinase that phosphorylates an RNA polymerase II-derived peptide. Biochem Biophys Res Commun. 1989 Mar 15;159(2):508–515. doi: 10.1016/0006-291x(89)90022-3. [DOI] [PubMed] [Google Scholar]
  42. Tamm I. Definition of subclasses of nucleoplasmic RNA. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5011–5015. doi: 10.1073/pnas.74.11.5011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Watson R. J. A transcriptional arrest mechanism involved in controlling constitutive levels of mouse c-myb mRNA. Oncogene. 1988 Mar;2(3):267–272. [PubMed] [Google Scholar]
  44. Watson R. J. Expression of the c-myb and c-myc genes is regulated independently in differentiating mouse erythroleukemia cells by common processes of premature transcription arrest and increased mRNA turnover. Mol Cell Biol. 1988 Sep;8(9):3938–3942. doi: 10.1128/mcb.8.9.3938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. 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]
  46. Zandomeni R., Bunick D., Ackerman S., Mittleman B., Weinmann R. Mechanism of action of DRB. III. Effect on specific in vitro initiation of transcription. J Mol Biol. 1983 Jul 5;167(3):561–574. doi: 10.1016/s0022-2836(83)80098-9. [DOI] [PubMed] [Google Scholar]
  47. Zandomeni R., Mittleman B., Bunick D., Ackerman S., Weinmann R. Mechanism of action of dichloro-beta-D-ribofuranosylbenzimidazole: effect on in vitro transcription. Proc Natl Acad Sci U S A. 1982 May;79(10):3167–3170. doi: 10.1073/pnas.79.10.3167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Zandomeni R., Zandomeni M. C., Shugar D., Weinmann R. Casein kinase type II is involved in the inhibition by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole of specific RNA polymerase II transcription. J Biol Chem. 1986 Mar 5;261(7):3414–3419. [PubMed] [Google Scholar]
  49. de Vegvar H. E., Lund E., Dahlberg J. E. 3' end formation of U1 snRNA precursors is coupled to transcription from snRNA promoters. Cell. 1986 Oct 24;47(2):259–266. doi: 10.1016/0092-8674(86)90448-4. [DOI] [PubMed] [Google Scholar]

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

RESOURCES