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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1989 Oct;86(19):7326–7330. doi: 10.1073/pnas.86.19.7326

Differential binding of nuclear factors to the intron 1 sequences containing the transcriptional pause site correlates with c-myb expression.

C D Reddy 1, E P Reddy 1
PMCID: PMC298054  PMID: 2678098

Abstract

The molecular mechanisms that modulate c-myb mRNA levels in hematopoietic cells appear to involve premature termination of transcription in the first intron of the gene. We have examined the DNA-protein interactions within the first intron of the c-myb gene and identified a 1.0-kilobase region that could be responsible for its transcriptional regulation. Using the mobility-shift assay, we show a direct correlation between the extent of sequence-specific protein binding to intron 1 DNA fragments, and c-myb mRNA levels in different cell types. During dimethyl sulfoxide-induced differentiation of mouse erythroleukemic cells, there was a dramatic decrease in these nuclear factors that correlated with the decrease in the levels of c-myb mRNA. Nucleotide sequence analysis and DNase I footprinting revealed the presence of putative regulatory elements that are implicated in the binding of these nuclear factors. We propose that binding of nuclear factors to the site of transcriptional pause could play an important role in the regulation of c-myb transcription.

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Selected References

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  1. BALUDA M. A., GOETZ I. E. Morphological conversion of cell cultures by avian myeloblastosis virus. Virology. 1961 Oct;15:185–199. doi: 10.1016/0042-6822(61)90234-3. [DOI] [PubMed] [Google Scholar]
  2. 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]
  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. Boyle W. J., Lipsick J. S., Baluda M. A. Antibodies to the evolutionarily conserved amino-terminal region of the v-myb-encoded protein detect the c-myb protein in widely divergent metazoan species. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4685–4689. doi: 10.1073/pnas.83.13.4685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  7. Craig R. W., Bloch A. Early decline in c-myb oncogene expression in the differentiation of human myeloblastic leukemia (ML-1) cells induced with 12-O-tetradecanoylphorbol-13-acetate. Cancer Res. 1984 Feb;44(2):442–446. [PubMed] [Google Scholar]
  8. 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]
  9. Duprey S. P., Boettiger D. Developmental regulation of c-myb in normal myeloid progenitor cells. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6937–6941. doi: 10.1073/pnas.82.20.6937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dynan W. S., Tjian R. Isolation of transcription factors that discriminate between different promoters recognized by RNA polymerase II. Cell. 1983 Mar;32(3):669–680. doi: 10.1016/0092-8674(83)90053-3. [DOI] [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. 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]
  13. Gonda T. J., Metcalf D. Expression of myb, myc and fos proto-oncogenes during the differentiation of a murine myeloid leukaemia. Nature. 1984 Jul 19;310(5974):249–251. doi: 10.1038/310249a0. [DOI] [PubMed] [Google Scholar]
  14. Gonda T. J., Sheiness D. K., Bishop J. M. Transcripts from the cellular homologs of retroviral oncogenes: distribution among chicken tissues. Mol Cell Biol. 1982 Jun;2(6):617–624. doi: 10.1128/mcb.2.6.617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hoey T., Levine M. Divergent homeo box proteins recognize similar DNA sequences in Drosophila. Nature. 1988 Apr 28;332(6167):858–861. doi: 10.1038/332858a0. [DOI] [PubMed] [Google Scholar]
  16. Klempnauer K. H., Bonifer C., Sippel A. E. Identification and characterization of the protein encoded by the human c-myb proto-oncogene. EMBO J. 1986 Aug;5(8):1903–1911. doi: 10.1002/j.1460-2075.1986.tb04443.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Klinken S. P., Holmes K. L., Morse H. C., 3rd, Thorgeirsson S. S. Transcriptional and post-transcriptional regulation of c-myc, c-myb, and p53 during proliferation and differentiation of murine erythroleukemia cells treated with DFMO and DMSO. Exp Cell Res. 1988 Oct;178(2):185–198. doi: 10.1016/0014-4827(88)90390-4. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Lachman H. M., Skoultchi A. I. Expression of c-myc changes during differentiation of mouse erythroleukaemia cells. Nature. 1984 Aug 16;310(5978):592–594. doi: 10.1038/310592a0. [DOI] [PubMed] [Google Scholar]
  20. Lavu S., Reddy E. P. Structural organization and nucleotide sequence of mouse c-myb oncogene: activation in ABPL tumors is due to viral integration in an intron which results in the deletion of the 5' coding sequences. Nucleic Acids Res. 1986 Jul 11;14(13):5309–5320. doi: 10.1093/nar/14.13.5309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lee W., Mitchell P., Tjian R. Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements. Cell. 1987 Jun 19;49(6):741–752. doi: 10.1016/0092-8674(87)90612-x. [DOI] [PubMed] [Google Scholar]
  22. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Pauza C. D. Regulation of human T-lymphocyte gene expression by interleukin 2: immediate-response genes include the proto-oncogene c-myb. Mol Cell Biol. 1987 Jan;7(1):342–348. doi: 10.1128/mcb.7.1.342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ramsay R. G., Ikeda K., Rifkind R. A., Marks P. A. Changes in gene expression associated with induced differentiation of erythroleukemia: protooncogenes, globin genes, and cell division. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6849–6853. doi: 10.1073/pnas.83.18.6849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rosson D., Dugan D., Reddy E. P. Aberrant splicing events that are induced by proviral integration: implications for myb oncogene activation. Proc Natl Acad Sci U S A. 1987 May;84(10):3171–3175. doi: 10.1073/pnas.84.10.3171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Roussel M., Saule S., Lagrou C., Rommens C., Beug H., Graf T., Stehelin D. Three new types of viral oncogene of cellular origin specific for haematopoietic cell transformation. Nature. 1979 Oct 11;281(5731):452–455. doi: 10.1038/281452a0. [DOI] [PubMed] [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Singh H., Sen R., Baltimore D., Sharp P. A. A nuclear factor that binds to a conserved sequence motif in transcriptional control elements of immunoglobulin genes. Nature. 1986 Jan 9;319(6049):154–158. doi: 10.1038/319154a0. [DOI] [PubMed] [Google Scholar]
  30. Staudt L. M., Singh H., Sen R., Wirth T., Sharp P. A., Baltimore D. A lymphoid-specific protein binding to the octamer motif of immunoglobulin genes. Nature. 1986 Oct 16;323(6089):640–643. doi: 10.1038/323640a0. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. 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]
  34. Westin E. H., Gallo R. C., Arya S. K., Eva A., Souza L. M., Baluda M. A., Aaronson S. A., Wong-Staal F. Differential expression of the amv gene in human hematopoietic cells. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2194–2198. doi: 10.1073/pnas.79.7.2194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wingrove T. G., Watt R., Keng P., Macara I. G. Stabilization of myc proto-oncogene proteins during Friend murine erythroleukemia cell differentiation. J Biol Chem. 1988 Jun 25;263(18):8918–8924. [PubMed] [Google Scholar]
  36. Yuen L., Davison A. J., Moss B. Early promoter-binding factor from vaccinia virions. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6069–6073. doi: 10.1073/pnas.84.17.6069. [DOI] [PMC free article] [PubMed] [Google Scholar]

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