Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1993 Aug 25;21(17):3935–3942. doi: 10.1093/nar/21.17.3935

Purification by DNA affinity precipitation of the cellular factors HEB1-p67 and HEB1-p94 which bind specifically to the human T-cell leukemia virus type-I 21 bp enhancer.

G Lombard-Platet 1, P Jalinot 1
PMCID: PMC309971  PMID: 8371970

Abstract

Transcription driven by the proviral promoter of the Human T-cell Leukemia Virus type I (HTLV-I) is tightly regulated by the Tax1 transactivator. This viral protein potently induces the enhancer activity of a 21 bp motif repeated three times in the promoter. We have previously shown that this induction results from the binding of Tax1 to this enhancer sequence and that this association is mediated by the cellular factor HEB1. In this paper we report the purification of this factor by chromatography and DNA affinity precipitation. The latter method allowed a rapid and efficient purification which led to the identification of two polypeptides with molecular masses of 94- and 67-kDa, named HEB1-p94 and HEB1-p67, respectively. DMS methylation interference and UV crosslinking experiments indicated that both proteins formed different nucleo-protein complexes, but had the same DNA specificity. Study of the interaction of these two proteins with Tax1 showed that only HEB1-p67 can specifically interact with Tax1.

Full text

PDF
3935

Images in this article

3937Image
on p.3937
3938Image
on p.3938
3938Image
on p.3938
3939Image
on p.3939
3940Image
on p.3940
3940Image
on p.3940

Selected References

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

  1. Alexandre C., Verrier B. Four regulatory elements in the human c-fos promoter mediate transactivation by HTLV-1 Tax protein. Oncogene. 1991 Apr;6(4):543–551. [PubMed] [Google Scholar]
  2. Aoyama N., Nagase T., Sawazaki T., Mizuguchi G., Nakagoshi H., Fujisawa J. I., Yoshida M., Ishii S. Overlap of the p53-responsive element and cAMP-responsive element in the enhancer of human T-cell leukemia virus type I. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5403–5407. doi: 10.1073/pnas.89.12.5403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ballard D. W., Böhnlein E., Lowenthal J. W., Wano Y., Franza B. R., Greene W. C. HTLV-I tax induces cellular proteins that activate the kappa B element in the IL-2 receptor alpha gene. Science. 1988 Sep 23;241(4873):1652–1655. doi: 10.1126/science.241.4873.1652. [DOI] [PubMed] [Google Scholar]
  4. Brady J., Jeang K. T., Duvall J., Khoury G. Identification of p40x-responsive regulatory sequences within the human T-cell leukemia virus type I long terminal repeat. J Virol. 1987 Jul;61(7):2175–2181. doi: 10.1128/jvi.61.7.2175-2181.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Béraud C., Lombard-Platet G., Michal Y., Jalinot P. Binding of the HTLV-I Tax1 transactivator to the inducible 21 bp enhancer is mediated by the cellular factor HEB1. EMBO J. 1991 Dec;10(12):3795–3803. doi: 10.1002/j.1460-2075.1991.tb04949.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crenon I., Béraud C., Simard P., Montagne J., Veschambre P., Jalinot P. The transcriptionally active factors mediating the effect of the HTLV-I Tax transactivator on the IL-2R alpha kappa B enhancer include the product of the c-rel proto-oncogene. Oncogene. 1993 Apr;8(4):867–875. [PubMed] [Google Scholar]
  7. Fujii M., Tsuchiya H., Chuhjo T., Akizawa T., Seiki M. Interaction of HTLV-1 Tax1 with p67SRF causes the aberrant induction of cellular immediate early genes through CArG boxes. Genes Dev. 1992 Nov;6(11):2066–2076. doi: 10.1101/gad.6.11.2066. [DOI] [PubMed] [Google Scholar]
  8. Fujisawa J., Toita M., Yoshida M. A unique enhancer element for the trans activator (p40tax) of human T-cell leukemia virus type I that is distinct from cyclic AMP- and 12-O-tetradecanoylphorbol-13-acetate-responsive elements. J Virol. 1989 Aug;63(8):3234–3239. doi: 10.1128/jvi.63.8.3234-3239.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fujisawa J., Toita M., Yoshimura T., Yoshida M. The indirect association of human T-cell leukemia virus tax protein with DNA results in transcriptional activation. J Virol. 1991 Aug;65(8):4525–4528. doi: 10.1128/jvi.65.8.4525-4528.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gonzalez G. A., Montminy M. R. Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133. Cell. 1989 Nov 17;59(4):675–680. doi: 10.1016/0092-8674(89)90013-5. [DOI] [PubMed] [Google Scholar]
  11. Jackson S. P., Tjian R. O-glycosylation of eukaryotic transcription factors: implications for mechanisms of transcriptional regulation. Cell. 1988 Oct 7;55(1):125–133. doi: 10.1016/0092-8674(88)90015-3. [DOI] [PubMed] [Google Scholar]
  12. Kumar G., Sharma A. K., Jayaraman K. Incorporation of BrdU in a DNA fragment may affect protein-DNA interactions in a site-dependent manner. Oncogene. 1992 Jul;7(7):1453–1455. [PubMed] [Google Scholar]
  13. Leung K., Nabel G. J. HTLV-1 transactivator induces interleukin-2 receptor expression through an NF-kappa B-like factor. Nature. 1988 Jun 23;333(6175):776–778. doi: 10.1038/333776a0. [DOI] [PubMed] [Google Scholar]
  14. Marriott S. J., Boros I., Duvall J. F., Brady J. N. Indirect binding of human T-cell leukemia virus type I tax1 to a responsive element in the viral long terminal repeat. Mol Cell Biol. 1989 Oct;9(10):4152–4160. doi: 10.1128/mcb.9.10.4152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Marriott S. J., Lindholm P. F., Brown K. M., Gitlin S. D., Duvall J. F., Radonovich M. F., Brady J. N. A 36-kilodalton cellular transcription factor mediates an indirect interaction of human T-cell leukemia/lymphoma virus type I TAX1 with a responsive element in the viral long terminal repeat. Mol Cell Biol. 1990 Aug;10(8):4192–4201. doi: 10.1128/mcb.10.8.4192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Montagne J., Béraud C., Crenon I., Lombard-Platet G., Gazzolo L., Sergeant A., Jalinot P. Tax1 induction of the HTLV-I 21 bp enhancer requires cooperation between two cellular DNA-binding proteins. EMBO J. 1990 Mar;9(3):957–964. doi: 10.1002/j.1460-2075.1990.tb08194.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Muchardt C., Seeler J. S., Nirula A., Gong S., Gaynor R. Transcription factor AP-2 activates gene expression of HTLV-I. EMBO J. 1992 Jul;11(7):2573–2581. doi: 10.1002/j.1460-2075.1992.tb05322.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nakamura M., Niki M., Ohtani K., Sugamura K. Differential activation of the 21-base-pair enhancer element of human T-cell leukemia virus type I by its own trans-activator and cyclic AMP. Nucleic Acids Res. 1989 Jul 11;17(13):5207–5221. doi: 10.1093/nar/17.13.5207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nyborg J. K., Dynan W. S. Interaction of cellular proteins with the human T-cell leukemia virus type I transcriptional control region. Purification of cellular proteins that bind the 21-base pair repeat elements. J Biol Chem. 1990 May 15;265(14):8230–8236. [PubMed] [Google Scholar]
  20. Park R. E., Haseltine W. A., Rosen C. A. A nuclear factor is required for transactivation of HTLV-I gene expression. Oncogene. 1988 Sep;3(3):275–279. [PubMed] [Google Scholar]
  21. Ruben S., Poteat H., Tan T. H., Kawakami K., Roeder R., Haseltine W., Rosen C. A. Cellular transcription factors and regulation of IL-2 receptor gene expression by HTLV-I tax gene product. Science. 1988 Jul 1;241(4861):89–92. doi: 10.1126/science.2838905. [DOI] [PubMed] [Google Scholar]
  22. Suzuki T., Fujisawa J. I., Toita M., Yoshida M. The trans-activator tax of human T-cell leukemia virus type 1 (HTLV-1) interacts with cAMP-responsive element (CRE) binding and CRE modulator proteins that bind to the 21-base-pair enhancer of HTLV-1. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):610–614. doi: 10.1073/pnas.90.2.610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tan T. H., Horikoshi M., Roeder R. G. Purification and characterization of multiple nuclear factors that bind to the TAX-inducible enhancer within the human T-cell leukemia virus type 1 long terminal repeat. Mol Cell Biol. 1989 Apr;9(4):1733–1745. doi: 10.1128/mcb.9.4.1733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tsujimoto A., Nyunoya H., Morita T., Sato T., Shimotohno K. Isolation of cDNAs for DNA-binding proteins which specifically bind to a tax-responsive enhancer element in the long terminal repeat of human T-cell leukemia virus type I. J Virol. 1991 Mar;65(3):1420–1426. doi: 10.1128/jvi.65.3.1420-1426.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yoshimura T., Fujisawa J., Yoshida M. Multiple cDNA clones encoding nuclear proteins that bind to the tax-dependent enhancer of HTLV-1: all contain a leucine zipper structure and basic amino acid domain. EMBO J. 1990 Aug;9(8):2537–2542. doi: 10.1002/j.1460-2075.1990.tb07434.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zhao L. J., Giam C. Z. Human T-cell lymphotropic virus type I (HTLV-I) transcriptional activator, Tax, enhances CREB binding to HTLV-I 21-base-pair repeats by protein-protein interaction. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):7070–7074. doi: 10.1073/pnas.89.15.7070. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES