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. 1991 May;11(5):2832–2841. doi: 10.1128/mcb.11.5.2832

Sequence requirements for premature transcription arrest within the first intron of the mouse c-fos gene.

N Mechti 1, M Piechaczyk 1, J M Blanchard 1, P Jeanteur 1, B Lebleu 1
PMCID: PMC360067  PMID: 1901950

Abstract

A strong block to the elongation of nascent RNA transcripts by RNA polymerase II occurs in the 5' part of the mammalian c-fos proto-oncogene. In addition to the control of initiation, this mechanism contributes to transcriptional regulation of the gene. In vitro transcription experiments using nuclear extracts and purified transcription templates allowed us to map a unique arrest site within the mouse first intron 385 nucleotides downstream from the promoter. This position is in keeping with that estimated from nuclear run-on assays performed with short DNA probes and thus suggests that it corresponds to the actual block in vivo. Moreover, we have shown that neither the c-fos promoter nor upstream sequences are absolute requirements for an efficient transcription arrest both in vivo and in vitro. Finally, we have characterized a 103-nucleotide-long intron 1 motif comprising the arrest site and sufficient for obtaining the block in a cell-free transcription assay.

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Selected References

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

  1. 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]
  2. 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]
  3. 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]
  4. Blanchard J. M., Piechaczyk M., Fort P., Bonnieu A., Mechti N., Rech J., Cuny M., Lebleu B., Jeanteur P. The regulatory strategies of c-myc and c-fos proto-oncogenes share some common mechanisms. Biochimie. 1988 Jul;70(7):877–884. doi: 10.1016/0300-9084(88)90228-3. [DOI] [PubMed] [Google Scholar]
  5. Bonnieu A., Rech J., Jeanteur P., Fort P. Requirements for c-fos mRNA down regulation in growth stimulated murine cells. Oncogene. 1989 Jul;4(7):881–888. [PubMed] [Google Scholar]
  6. 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]
  7. Chiu R., Boyle W. J., Meek J., Smeal T., Hunter T., Karin M. The c-Fos protein interacts with c-Jun/AP-1 to stimulate transcription of AP-1 responsive genes. Cell. 1988 Aug 12;54(4):541–552. doi: 10.1016/0092-8674(88)90076-1. [DOI] [PubMed] [Google Scholar]
  8. Chung J., Sussman D. J., Zeller R., Leder P. The c-myc gene encodes superimposed RNA polymerase II and III promoters. Cell. 1987 Dec 24;51(6):1001–1008. doi: 10.1016/0092-8674(87)90586-1. [DOI] [PubMed] [Google Scholar]
  9. Ciliberto G., Dathan N., Frank R., Philipson L., Mattaj I. W. Formation of the 3' end on U snRNAs requires at least three sequence elements. EMBO J. 1986 Nov;5(11):2931–2937. doi: 10.1002/j.1460-2075.1986.tb04589.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Collart M. A., Tourkine N., Belin D., Vassalli P., Jeanteur P., Blanchard J. M. c-fos gene transcription in murine macrophages is modulated by a calcium-dependent block to elongation in intron 1. Mol Cell Biol. 1991 May;11(5):2826–2831. doi: 10.1128/mcb.11.5.2826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  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. Franza B. R., Jr, Rauscher F. J., 3rd, Josephs S. F., Curran T. The Fos complex and Fos-related antigens recognize sequence elements that contain AP-1 binding sites. Science. 1988 Mar 4;239(4844):1150–1153. doi: 10.1126/science.2964084. [DOI] [PubMed] [Google Scholar]
  15. Gilman M. Z. The c-fos serum response element responds to protein kinase C-dependent and -independent signals but not to cyclic AMP. Genes Dev. 1988 Apr;2(4):394–402. doi: 10.1101/gad.2.4.394. [DOI] [PubMed] [Google Scholar]
  16. Hagenbüchle O., Wellauer P. K., Cribbs D. L., Schibler U. Termination of transcription in the mouse alpha-amylase gene Amy-2a occurs at multiple sites downstream of the polyadenylation site. Cell. 1984 Oct;38(3):737–744. doi: 10.1016/0092-8674(84)90269-1. [DOI] [PubMed] [Google Scholar]
  17. Halazonetis T. D., Georgopoulos K., Greenberg M. E., Leder P. c-Jun dimerizes with itself and with c-Fos, forming complexes of different DNA binding affinities. Cell. 1988 Dec 2;55(5):917–924. doi: 10.1016/0092-8674(88)90147-x. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Hofer E., Darnell J. E., Jr The primary transcription unit of the mouse beta-major globin gene. Cell. 1981 Feb;23(2):585–593. doi: 10.1016/0092-8674(81)90154-9. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Lamb N. J., Fernandez A., Tourkine N., Jeanteur P., Blanchard J. M. Demonstration in living cells of an intragenic negative regulatory element within the rodent c-fos gene. Cell. 1990 May 4;61(3):485–496. doi: 10.1016/0092-8674(90)90530-r. [DOI] [PubMed] [Google Scholar]
  23. Lattier D. L., States J. C., Hutton J. J., Wiginton D. A. Cell type-specific transcriptional regulation of the human adenosine deaminase gene. Nucleic Acids Res. 1989 Feb 11;17(3):1061–1076. doi: 10.1093/nar/17.3.1061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mechti N., Piechaczyk M., Blanchard J. M., Marty L., Bonnieu A., Jeanteur P., Lebleu B. Transcriptional and post-transcriptional regulation of c-myc expression during the differentiation of murine erythroleukemia Friend cells. Nucleic Acids Res. 1986 Dec 22;14(24):9653–9666. doi: 10.1093/nar/14.24.9653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Miyamoto N. G., Moncollin V., Wintzerith M., Hen R., Egly J. M., Chambon P. Stimulation of in vitro transcription by the upstream element of the adenovirus-2 major late promoter involves a specific factor. Nucleic Acids Res. 1984 Dec 11;12(23):8779–8799. doi: 10.1093/nar/12.23.8779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mok M., Maderious A., Chen-Kiang S. Premature termination by human RNA polymerase II occurs temporally in the adenovirus major late transcriptional unit. Mol Cell Biol. 1984 Oct;4(10):2031–2040. doi: 10.1128/mcb.4.10.2031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nakabeppu Y., Ryder K., Nathans D. DNA binding activities of three murine Jun proteins: stimulation by Fos. Cell. 1988 Dec 2;55(5):907–915. doi: 10.1016/0092-8674(88)90146-8. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Proudfoot N. J. How RNA polymerase II terminates transcription in higher eukaryotes. Trends Biochem Sci. 1989 Mar;14(3):105–110. doi: 10.1016/0968-0004(89)90132-1. [DOI] [PubMed] [Google Scholar]
  30. Ramamurthy V., Maa M. C., Harless M. L., Wright D. A., Kellems R. E. Sequence requirements for transcriptional arrest in exon 1 of the murine adenosine deaminase gene. Mol Cell Biol. 1990 Apr;10(4):1484–1491. doi: 10.1128/mcb.10.4.1484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rauscher F. J., 3rd, Cohen D. R., Curran T., Bos T. J., Vogt P. K., Bohmann D., Tjian R., Franza B. R., Jr Fos-associated protein p39 is the product of the jun proto-oncogene. Science. 1988 May 20;240(4855):1010–1016. doi: 10.1126/science.3130660. [DOI] [PubMed] [Google Scholar]
  32. Re G. G., Antoun G. R., Zipf T. F. Modulation of a constitutive transcriptional block at exon-1 controls human c-myc oncogene expression. Oncogene. 1990 Aug;5(8):1247–1250. [PubMed] [Google Scholar]
  33. Rech J., Fort P. Versatile vectors for pulsed expression in eukaryotic cells. Nucleic Acids Res. 1989 Apr 11;17(7):2874–2874. doi: 10.1093/nar/17.7.2874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Reddy C. D., Reddy E. P. Differential binding of nuclear factors to the intron 1 sequences containing the transcriptional pause site correlates with c-myb expression. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7326–7330. doi: 10.1073/pnas.86.19.7326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Resnekov O., Aloni Y. RNA polymerase II is capable of pausing and prematurely terminating transcription at a precise location in vivo and in vitro. Proc Natl Acad Sci U S A. 1989 Jan;86(1):12–16. doi: 10.1073/pnas.86.1.12. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rüther U., Wagner E. F., Müller R. Analysis of the differentiation-promoting potential of inducible c-fos genes introduced into embryonal carcinoma cells. EMBO J. 1985 Jul;4(7):1775–1781. doi: 10.1002/j.1460-2075.1985.tb03850.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sassone-Corsi P., Lamph W. W., Kamps M., Verma I. M. fos-associated cellular p39 is related to nuclear transcription factor AP-1. Cell. 1988 Aug 12;54(4):553–560. doi: 10.1016/0092-8674(88)90077-3. [DOI] [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. Sobczak J., Mechti N., Tournier M. F., Blanchard J. M., Duguet M. c-myc and c-fos gene regulation during mouse liver regeneration. Oncogene. 1989 Dec;4(12):1503–1508. [PubMed] [Google Scholar]
  40. Southgate C., Zapp M. L., Green M. R. Activation of transcription by HIV-1 Tat protein tethered to nascent RNA through another protein. Nature. 1990 Jun 14;345(6276):640–642. doi: 10.1038/345640a0. [DOI] [PubMed] [Google Scholar]
  41. Spencer C. A., Groudine M. Transcription elongation and eukaryotic gene regulation. Oncogene. 1990 Jun;5(6):777–785. [PubMed] [Google Scholar]
  42. 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]
  43. Treisman R. Identification of a protein-binding site that mediates transcriptional response of the c-fos gene to serum factors. Cell. 1986 Aug 15;46(4):567–574. doi: 10.1016/0092-8674(86)90882-2. [DOI] [PubMed] [Google Scholar]
  44. Treisman R. Transient accumulation of c-fos RNA following serum stimulation requires a conserved 5' element and c-fos 3' sequences. Cell. 1985 Oct;42(3):889–902. doi: 10.1016/0092-8674(85)90285-5. [DOI] [PubMed] [Google Scholar]
  45. 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]
  46. 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]
  47. Zuker M., Stiegler P. Optimal computer folding of large RNA sequences using thermodynamics and auxiliary information. Nucleic Acids Res. 1981 Jan 10;9(1):133–148. doi: 10.1093/nar/9.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]

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