Abstract
A block to c-myc transcription elongation has been observed in Xenopus oocytes and mammalian cells. Here, we show that the distribution of RNA polymerase II transcription complexes in the c-myc promoter proximal region in Xenopus oocytes is different from that observed previously in mammalian cells. Thus, there are major differences in the c-myc elongation block observed in the two systems. In addition, as first reported for a Xenopus tubulin gene (K. M. Middleton and G. T. Morgan, Mol. Cell. Biol. 10:727-735, 1990). c-myc template titration experiments reveal the existence of two classes of RNA polymerase II transcription complexes in oocytes: one (at low template concentration) that is capable of reading through downstream sites of premature termination, and another (high template concentration) that does not. We show that these classes of polymerases are distinct from those previously identified by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), which distinguishes transcription complexes on the basis of transcribed distance, rather than on the basis of differential elongation through sites of premature termination. We also show that mutations that affect the efficiency of initiation of transcription from the c-myc P2 promoter can influence premature termination by at least two mechanisms: TATA box mutations function by the titration effect (decrease in transcription initiation results in a relative decrease in premature termination), while an upstream activator (E2F) site functions by contributing to the assembly of polymerase complexes competent to traverse the downstream sites of premature termination.
Full text
PDF











Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ashfield R., Enriquez-Harris P., Proudfoot N. J. Transcriptional termination between the closely linked human complement genes C2 and factor B: common termination factor for C2 and c-myc? EMBO J. 1991 Dec;10(13):4197–4207. doi: 10.1002/j.1460-2075.1991.tb04998.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Asselin C., Nepveu A., Marcu K. B. Molecular requirements for transcriptional initiation of the murine c-myc gene. Oncogene. 1989 May;4(5):549–558. [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- Bossone S. A., Asselin C., Patel A. J., Marcu K. B. MAZ, a zinc finger protein, binds to c-MYC and C2 gene sequences regulating transcriptional initiation and termination. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7452–7456. doi: 10.1073/pnas.89.16.7452. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Buckler A. J., Rothstein T. L., Sonenshein G. E. Transcriptional control of c-myc gene expression during stimulation of murine B lymphocytes. J Immunol. 1990 Jul 15;145(2):732–736. [PubMed] [Google Scholar]
- Burton Z. F., Killeen M., Sopta M., Ortolan L. G., Greenblatt J. RAP30/74: a general initiation factor that binds to RNA polymerase II. Mol Cell Biol. 1988 Apr;8(4):1602–1613. doi: 10.1128/mcb.8.4.1602. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chafin D. R., Claussen T. J., Price D. H. Identification and purification of a yeast protein that affects elongation by RNA polymerase II. J Biol Chem. 1991 May 15;266(14):9256–9262. [PubMed] [Google Scholar]
- Chang Y., Spicer D. B., Sonenshein G. E. Effects of IL-3 on promoter usage, attenuation and antisense transcription of the c-myc oncogene in the IL-3-dependent Ba/F3 early pre-B cell line. Oncogene. 1991 Nov;6(11):1979–1982. [PubMed] [Google Scholar]
- 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]
- 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]
- Eick D., Berger R., Polack A., Bornkamm G. W. Transcription of c-myc in human mononuclear cells is regulated by an elongation block. Oncogene. 1987;2(1):61–65. [PubMed] [Google Scholar]
- 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]
- Flores O., Maldonado E., Reinberg D. Factors involved in specific transcription by mammalian RNA polymerase II. Factors IIE and IIF independently interact with RNA polymerase II. J Biol Chem. 1989 May 25;264(15):8913–8921. [PubMed] [Google Scholar]
- 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]
- Hiebert S. W., Lipp M., Nevins J. R. E1A-dependent trans-activation of the human MYC promoter is mediated by the E2F factor. Proc Natl Acad Sci U S A. 1989 May;86(10):3594–3598. doi: 10.1073/pnas.86.10.3594. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Innis J. W., Kellems R. E. A heat-labile factor promotes premature 3' end formation in exon 1 of the murine adenosine deaminase gene in a cell-free transcription system. Mol Cell Biol. 1991 Nov;11(11):5398–5409. doi: 10.1128/mcb.11.11.5398. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Kato H., Sumimoto H., Pognonec P., Chen C. H., Rosen C. A., Roeder R. G. HIV-1 Tat acts as a processivity factor in vitro in conjunction with cellular elongation factors. Genes Dev. 1992 Apr;6(4):655–666. doi: 10.1101/gad.6.4.655. [DOI] [PubMed] [Google Scholar]
- Kephart D. D., Marshall N. F., Price D. H. Stability of Drosophila RNA polymerase II elongation complexes in vitro. Mol Cell Biol. 1992 May;12(5):2067–2077. doi: 10.1128/mcb.12.5.2067. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Krauskopf A., Bengal E., Aloni Y. The block to transcription elongation at the minute virus of mice attenuator is regulated by cellular elongation factors. Mol Cell Biol. 1991 Jul;11(7):3515–3521. doi: 10.1128/mcb.11.7.3515. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Lee H., Kraus K. W., Wolfner M. F., Lis J. T. DNA sequence requirements for generating paused polymerase at the start of hsp70. Genes Dev. 1992 Feb;6(2):284–295. doi: 10.1101/gad.6.2.284. [DOI] [PubMed] [Google Scholar]
- Lindsten T., June C. H., Thompson C. B. Multiple mechanisms regulate c-myc gene expression during normal T cell activation. EMBO J. 1988 Sep;7(9):2787–2794. doi: 10.1002/j.1460-2075.1988.tb03133.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Lüscher B., Eisenman R. N. New light on Myc and Myb. Part I. Myc. Genes Dev. 1990 Dec;4(12A):2025–2035. doi: 10.1101/gad.4.12a.2025. [DOI] [PubMed] [Google Scholar]
- Marciniak R. A., Sharp P. A. HIV-1 Tat protein promotes formation of more-processive elongation complexes. EMBO J. 1991 Dec;10(13):4189–4196. doi: 10.1002/j.1460-2075.1991.tb04997.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marshall N. F., Price D. H. Control of formation of two distinct classes of RNA polymerase II elongation complexes. Mol Cell Biol. 1992 May;12(5):2078–2090. doi: 10.1128/mcb.12.5.2078. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Meulia T., Krumm A., Spencer C., Groudine M. Sequences in the human c-myc P2 promoter affect the elongation and premature termination of transcripts initiated from the upstream P1 promoter. Mol Cell Biol. 1992 Oct;12(10):4590–4600. doi: 10.1128/mcb.12.10.4590. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Middleton K. M., Morgan G. T. Premature termination of transcription can be induced on an injected alpha-tubulin gene in Xenopus oocytes. Mol Cell Biol. 1990 Feb;10(2):727–735. doi: 10.1128/mcb.10.2.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller H., Asselin C., Dufort D., Yang J. Q., Gupta K., Marcu K. B., Nepveu A. A cis-acting element in the promoter region of the murine c-myc gene is necessary for transcriptional block. Mol Cell Biol. 1989 Dec;9(12):5340–5349. doi: 10.1128/mcb.9.12.5340. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mirkovitch J., Darnell J. E., Jr Mapping of RNA polymerase on mammalian genes in cells and nuclei. Mol Biol Cell. 1992 Oct;3(10):1085–1094. doi: 10.1091/mbc.3.10.1085. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mukherjee R., Molloy G. R. 5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole inhibits transcription of the beta-hemoglobin gene in vivo at initiation. J Biol Chem. 1987 Oct 5;262(28):13697–13705. [PubMed] [Google Scholar]
- 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]
- O'Brien T., Lis J. T. RNA polymerase II pauses at the 5' end of the transcriptionally induced Drosophila hsp70 gene. Mol Cell Biol. 1991 Oct;11(10):5285–5290. doi: 10.1128/mcb.11.10.5285. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pyrc J. J., Moberg K. H., Hall D. J. Isolation of a novel cDNA encoding a zinc-finger protein that binds to two sites within the c-myc promoter. Biochemistry. 1992 Apr 28;31(16):4102–4110. doi: 10.1021/bi00131a029. [DOI] [PubMed] [Google Scholar]
- 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]
- Reines D., Chamberlin M. J., Kane C. M. Transcription elongation factor SII (TFIIS) enables RNA polymerase II to elongate through a block to transcription in a human gene in vitro. J Biol Chem. 1989 Jun 25;264(18):10799–10809. [PubMed] [Google Scholar]
- Reines D. Elongation factor-dependent transcript shortening by template-engaged RNA polymerase II. J Biol Chem. 1992 Feb 25;267(6):3795–3800. [PMC free article] [PubMed] [Google Scholar]
- Roberts S., Bentley D. L. Distinct modes of transcription read through or terminate at the c-myc attenuator. EMBO J. 1992 Mar;11(3):1085–1093. doi: 10.1002/j.1460-2075.1992.tb05147.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rougvie A. E., Lis J. T. The RNA polymerase II molecule at the 5' end of the uninduced hsp70 gene of D. melanogaster is transcriptionally engaged. Cell. 1988 Sep 9;54(6):795–804. doi: 10.1016/s0092-8674(88)91087-2. [DOI] [PubMed] [Google Scholar]
- SivaRaman L., Reines D., Kane C. M. Purified elongation factor SII is sufficient to promote read-through by purified RNA polymerase II at specific termination sites in the human histone H3.3 gene. J Biol Chem. 1990 Aug 25;265(24):14554–14560. [PubMed] [Google Scholar]
- 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]
- Spencer C. A., Groudine M. Control of c-myc regulation in normal and neoplastic cells. Adv Cancer Res. 1991;56:1–48. doi: 10.1016/s0065-230x(08)60476-5. [DOI] [PubMed] [Google Scholar]
- Spencer C. A., Groudine M. Transcription elongation and eukaryotic gene regulation. Oncogene. 1990 Jun;5(6):777–785. [PubMed] [Google Scholar]
- 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]
- 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]
- Strobl L. J., Eick D. Hold back of RNA polymerase II at the transcription start site mediates down-regulation of c-myc in vivo. EMBO J. 1992 Sep;11(9):3307–3314. doi: 10.1002/j.1460-2075.1992.tb05409.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thalmeier K., Synovzik H., Mertz R., Winnacker E. L., Lipp M. Nuclear factor E2F mediates basic transcription and trans-activation by E1a of the human MYC promoter. Genes Dev. 1989 Apr;3(4):527–536. doi: 10.1101/gad.3.4.527. [DOI] [PubMed] [Google Scholar]
- 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]
- Wright S., Mirels L. F., Calayag M. C., Bishop J. M. Premature termination of transcription from the P1 promoter of the mouse c-myc gene. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11383–11387. doi: 10.1073/pnas.88.24.11383. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yanofsky C. Transcription attenuation. J Biol Chem. 1988 Jan 15;263(2):609–612. [PubMed] [Google Scholar]
- ar-Rushdi A., Nishikura K., Erikson J., Watt R., Rovera G., Croce C. M. Differential expression of the translocated and the untranslocated c-myc oncogene in Burkitt lymphoma. Science. 1983 Oct 28;222(4622):390–393. doi: 10.1126/science.6414084. [DOI] [PubMed] [Google Scholar]