Abstract
To analyze regulation of the human T-cell leukemia virus type I (HTLV-I) long terminal repeat (LTR), cell lines were generated from LTR-tax x LTR-beta-galactosidase (beta-Gal) doubly transgenic mouse fibroblastic tumors. The HTLV-I LTR directs expression of both the tax and lacZ genes, and Tax up-modulates both promoters in primary cells. However, once cells were transformed by tax, beta-Gal but not tax expression was suppressed. Supertransformation of these cells with v-src suppressed both beta-Gal and tax expression. This suppression was reversed by treatment with the tyrosine kinase inhibitor herbimycin A or protein kinase A inhibitor H8. Electrophoretic mobility shift assays demonstrated augmented binding in the R but not U3 region. This binding was competitively inhibited by a high-affinity CREB oligodeoxynucleotide and super-shifted with a specific CREB antibody. Treatment of cells with the cyclic AMP analog dibutyryl cyclic AMP also transiently increased the R region binding dramatically. In vitro DNase I footprint analysis identified a protein-binding sequence in the R region which corresponded with suppression. However, this target sequence lacked a conventional CREB-binding site. A 70.5-kDa DNA-binding protein was partially purified by affinity chromatography, along with a 49-kDa protein which reacted with CREB-specific sera. These data demonstrate that HTLV-I LTR suppression is associated with CREB factor binding in the R region, probably by direct interaction with a 70.5-kDa protein, and provide a novel mechanism for maintenance of viral latency.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Akerblom I. E., Slater E. P., Beato M., Baxter J. D., Mellon P. L. Negative regulation by glucocorticoids through interference with a cAMP responsive enhancer. Science. 1988 Jul 15;241(4863):350–353. doi: 10.1126/science.2838908. [DOI] [PubMed] [Google Scholar]
- Andrisani O., Dixon J. E. Identification and purification of a novel 120-kDa protein that recognizes the cAMP-responsive element. J Biol Chem. 1990 Feb 25;265(6):3212–3218. [PubMed] [Google Scholar]
- Arya S. K. Human immunodeficiency virus type 2 (HIV-2) gene expression: downmodulation by sequence elements downstream of the transcriptional initiation site. AIDS Res Hum Retroviruses. 1991 Dec;7(12):1007–1014. doi: 10.1089/aid.1991.7.1007. [DOI] [PubMed] [Google Scholar]
- Badley J. E., Bishop G. A., St John T., Frelinger J. A. A simple, rapid method for the purification of poly A+ RNA. Biotechniques. 1988 Feb;6(2):114–116. [PubMed] [Google Scholar]
- Batchelor A. H., O'Hare P. Regulation and cell-type-specific activity of a promoter located upstream of the latency-associated transcript of herpes simplex virus type 1. J Virol. 1990 Jul;64(7):3269–3279. doi: 10.1128/jvi.64.7.3269-3279.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bosselut R., Duvall J. F., Gégonne A., Bailly M., Hémar A., Brady J., Ghysdael J. The product of the c-ets-1 proto-oncogene and the related Ets2 protein act as transcriptional activators of the long terminal repeat of human T cell leukemia virus HTLV-1. EMBO J. 1990 Oct;9(10):3137–3144. doi: 10.1002/j.1460-2075.1990.tb07511.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brott B. K., Decker S., O'Brien M. C., Jove R. Molecular features of the viral and cellular Src kinases involved in interactions with the GTPase-activating protein. Mol Cell Biol. 1991 Oct;11(10):5059–5067. doi: 10.1128/mcb.11.10.5059. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown D. A., Kondo K. L., Wong S. W., Diamond D. J. Characterization of nuclear protein binding to the interferon-gamma promoter in quiescent and activated human T cells. Eur J Immunol. 1992 Sep;22(9):2419–2428. doi: 10.1002/eji.1830220935. [DOI] [PubMed] [Google Scholar]
- Bushman W. A., Wilson L. K., Luttrell D. K., Moyers J. S., Parsons S. J. Overexpression of c-src enhances beta-adrenergic-induced cAMP accumulation. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7462–7466. doi: 10.1073/pnas.87.19.7462. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Copeland N. G., Zelenetz A. D., Copper G. M. Transformation by subgenomic fragments of Rous sarcoma virus DNA. Cell. 1980 Apr;19(4):863–870. doi: 10.1016/0092-8674(80)90077-x. [DOI] [PubMed] [Google Scholar]
- Farabaugh P. J., Vimaladithan A., Türkel S., Johnson R., Zhao H. Three downstream sites repress transcription of a Ty2 retrotransposon in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Apr;13(4):2081–2090. doi: 10.1128/mcb.13.4.2081. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Franchini G., Wong-Staal F., Gallo R. C. Human T-cell leukemia virus (HTLV-I) transcripts in fresh and cultured cells of patients with adult T-cell leukemia. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6207–6211. doi: 10.1073/pnas.81.19.6207. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Franklin A. A., Kubik M. F., Uittenbogaard M. N., Brauweiler A., Utaisincharoen P., Matthews M. A., Dynan W. S., Hoeffler J. P., Nyborg J. K. Transactivation by the human T-cell leukemia virus Tax protein is mediated through enhanced binding of activating transcription factor-2 (ATF-2) ATF-2 response and cAMP element-binding protein (CREB). J Biol Chem. 1993 Oct 5;268(28):21225–21231. [PubMed] [Google Scholar]
- 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]
- Garcia R., Parikh N. U., Saya H., Gallick G. E. Effect of herbimycin A on growth and pp60c-src activity in human colon tumor cell lines. Oncogene. 1991 Nov;6(11):1983–1989. [PubMed] [Google Scholar]
- Gartenhaus R. B., Wong-Staal F., Klotman M. E. The promoter of human T-cell leukemia virus type-I is repressed by the immediate-early gene region of human cytomegalovirus in primary blood lymphocytes. Blood. 1991 Dec 1;78(11):2956–2961. [PubMed] [Google Scholar]
- Gégonne A., Bosselut R., Bailly R. A., Ghysdael J. Synergistic activation of the HTLV1 LTR Ets-responsive region by transcription factors Ets1 and Sp1. EMBO J. 1993 Mar;12(3):1169–1178. doi: 10.1002/j.1460-2075.1993.tb05758.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Habener J. F. Cyclic AMP response element binding proteins: a cornucopia of transcription factors. Mol Endocrinol. 1990 Aug;4(8):1087–1094. doi: 10.1210/mend-4-8-1087. [DOI] [PubMed] [Google Scholar]
- Hansen M. B., Nielsen S. E., Berg K. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Methods. 1989 May 12;119(2):203–210. doi: 10.1016/0022-1759(89)90397-9. [DOI] [PubMed] [Google Scholar]
- Hemmings B. A. cAMP mediated proteolysis of the catalytic subunit of cAMP-dependent protein kinase. FEBS Lett. 1986 Feb 3;196(1):126–130. doi: 10.1016/0014-5793(86)80226-5. [DOI] [PubMed] [Google Scholar]
- Hidaka H., Inagaki M., Kawamoto S., Sasaki Y. Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry. 1984 Oct 9;23(21):5036–5041. doi: 10.1021/bi00316a032. [DOI] [PubMed] [Google Scholar]
- Hinrichs S. H., Nerenberg M., Reynolds R. K., Khoury G., Jay G. A transgenic mouse model for human neurofibromatosis. Science. 1987 Sep 11;237(4820):1340–1343. doi: 10.1126/science.2888191. [DOI] [PubMed] [Google Scholar]
- Ho D. Y. Herpes simplex virus latency: molecular aspects. Prog Med Virol. 1992;39:76–115. [PubMed] [Google Scholar]
- Kadonaga J. T., Tjian R. Affinity purification of sequence-specific DNA binding proteins. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5889–5893. doi: 10.1073/pnas.83.16.5889. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kashanchi F., Duvall J. F., Lindholm P. F., Radonovich M. F., Brady J. N. Sequences downstream of the RNA initiation site regulate human T-cell lymphotropic virus type I basal gene expression. J Virol. 1993 May;67(5):2894–2902. doi: 10.1128/jvi.67.5.2894-2902.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kitajima I., Shinohara T., Bilakovics J., Brown D. A., Xu X., Nerenberg M. Ablation of transplanted HTLV-I Tax-transformed tumors in mice by antisense inhibition of NF-kappa B. Science. 1992 Dec 11;258(5089):1792–1795. doi: 10.1126/science.1299224. [DOI] [PubMed] [Google Scholar]
- Kitajima I., Shinohara T., Minor T., Bibbs L., Bilakovics J., Nerenberg M. Human T-cell leukemia virus type I tax transformation is associated with increased uptake of oligodeoxynucleotides in vitro and in vivo. J Biol Chem. 1992 Dec 25;267(36):25881–25888. [PubMed] [Google Scholar]
- Kwast-Welfeld J., de Belle I., Walker P. R., Whitfield J. F., Sikorska M. Identification of a new cAMP response element-binding factor by southwestern blotting. J Biol Chem. 1993 Sep 15;268(26):19581–19585. [PubMed] [Google Scholar]
- Levinger L. F., Lautenberger J. A. Human protein binding to DNA sequences surrounding the human T-cell lymphotropic virus type-I long terminal repeat polyadenylation site. Eur J Biochem. 1987 Aug 3;166(3):519–526. doi: 10.1111/j.1432-1033.1987.tb13544.x. [DOI] [PubMed] [Google Scholar]
- Miyoshi I., Kubonishi I., Yoshimoto S., Akagi T., Ohtsuki Y., Shiraishi Y., Nagata K., Hinuma Y. Type C virus particles in a cord T-cell line derived by co-cultivating normal human cord leukocytes and human leukaemic T cells. Nature. 1981 Dec 24;294(5843):770–771. doi: 10.1038/294770a0. [DOI] [PubMed] [Google Scholar]
- Moyers J. S., Bouton A. H., Parsons S. J. The sites of phosphorylation by protein kinase C and an intact SH2 domain are required for the enhanced response to beta-adrenergic agonists in cells overexpressing c-src. Mol Cell Biol. 1993 Apr;13(4):2391–2400. doi: 10.1128/mcb.13.4.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Nakamura M., Ohtani K., Hinuma Y., Sugamura K. Functional mapping of the activity of the R region in the human T-cell leukemia virus type I long terminal repeat to increase gene expression. Virus Genes. 1989 Mar;2(2):147–155. doi: 10.1007/BF00315258. [DOI] [PubMed] [Google Scholar]
- Nerenberg M. I., Minor T., Price J., Ernst D. N., Shinohara T., Schwarz H. Transgenic thymocytes are refractory to transformation by the human T-cell leukemia virus type I tax gene. J Virol. 1991 Jun;65(6):3349–3353. doi: 10.1128/jvi.65.6.3349-3353.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nerenberg M. I., Wiley C. A. Degeneration of oxidative muscle fibers in HTLV-1 tax transgenic mice. Am J Pathol. 1989 Dec;135(6):1025–1033. [PMC free article] [PubMed] [Google Scholar]
- Nerenberg M., Hinrichs S. H., Reynolds R. K., Khoury G., Jay G. The tat gene of human T-lymphotropic virus type 1 induces mesenchymal tumors in transgenic mice. Science. 1987 Sep 11;237(4820):1324–1329. doi: 10.1126/science.2888190. [DOI] [PubMed] [Google Scholar]
- Nichols M., Weih F., Schmid W., DeVack C., Kowenz-Leutz E., Luckow B., Boshart M., Schütz G. Phosphorylation of CREB affects its binding to high and low affinity sites: implications for cAMP induced gene transcription. EMBO J. 1992 Sep;11(9):3337–3346. doi: 10.1002/j.1460-2075.1992.tb05412.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Niki M., Ohtani K., Nakamura M., Sugamura K. Multistep regulation of enhancer activity of the 21-base-pair element of human T-cell leukemia virus type I. J Virol. 1992 Jul;66(7):4348–4357. doi: 10.1128/jvi.66.7.4348-4357.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nyborg J. K., Matthews M. A., Yucel J., Walls L., Golde W. T., Dynan W. S., Wachsman W. Interaction of host cell proteins with the human T-cell leukemia virus type I transcriptional control region. II. A comprehensive map of protein-binding sites facilitates construction of a simple chimeric promoter responsive to the viral tax2 gene product. J Biol Chem. 1990 May 15;265(14):8237–8242. [PubMed] [Google Scholar]
- Olashaw N. E., Pledger W. J. Cellular mechanisms regulating proliferation. Adv Second Messenger Phosphoprotein Res. 1988;22:139–173. [PubMed] [Google Scholar]
- Paskalis H., Felber B. K., Pavlakis G. N. Cis-acting sequences responsible for the transcriptional activation of human T-cell leukemia virus type I constitute a conditional enhancer. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6558–6562. doi: 10.1073/pnas.83.17.6558. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pierani A., Pouponnot C., Calothy G. Transcriptional downregulation of the retina-specific QR1 gene by pp60v-src and identification of a novel v-src-responsive unit. Mol Cell Biol. 1993 Jun;13(6):3401–3414. doi: 10.1128/mcb.13.6.3401. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Qureshi S. A., Joseph C. K., Rim M., Maroney A., Foster D. A. v-Src activates both protein kinase C-dependent and independent signaling pathways in murine fibroblasts. Oncogene. 1991 Jun;6(6):995–999. [PubMed] [Google Scholar]
- Risse G., Jooss K., Neuberg M., Brüller H. J., Müller R. Asymmetrical recognition of the palindromic AP1 binding site (TRE) by Fos protein complexes. EMBO J. 1989 Dec 1;8(12):3825–3832. doi: 10.1002/j.1460-2075.1989.tb08560.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rodland K. D., Pribnow D., Lenormand P., Chen S. L., Magun B. E. Characterization of a unique enhancer element responsive to cyclic adenosine 3',5'-monophosphate and elevated calcium. Mol Endocrinol. 1993 Jun;7(6):787–796. doi: 10.1210/mend.7.6.8395655. [DOI] [PubMed] [Google Scholar]
- Rosen C. A., Sodroski J. G., Haseltine W. A. Location of cis-acting regulatory sequences in the human T-cell leukemia virus type I long terminal repeat. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6502–6506. doi: 10.1073/pnas.82.19.6502. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Saggioro D., Panozzo M., Chieco-Bianchi L. Human T-lymphotropic virus type I transcriptional regulation by methylation. Cancer Res. 1990 Aug 15;50(16):4968–4973. [PubMed] [Google Scholar]
- Salahuddin S. Z., Markham P. D., Wong-Staal F., Franchini G., Kalyanaraman V. S., Gallo R. C. Restricted expression of human T-cell leukemia--lymphoma virus (HTLV) in transformed human umbilical cord blood lymphocytes. Virology. 1983 Aug;129(1):51–64. doi: 10.1016/0042-6822(83)90395-1. [DOI] [PubMed] [Google Scholar]
- Shalloway D., Zelenetz A. D., Cooper G. M. Molecular cloning and characterization of the chicken gene homologous to the transforming gene of Rous sarcoma virus. Cell. 1981 May;24(2):531–541. doi: 10.1016/0092-8674(81)90344-5. [DOI] [PubMed] [Google Scholar]
- Stagg R. B., Fletcher W. H. The hormone-induced regulation of contact-dependent cell-cell communication by phosphorylation. Endocr Rev. 1990 May;11(2):302–325. doi: 10.1210/edrv-11-2-302. [DOI] [PubMed] [Google Scholar]
- Stauber C., Altschmied J., Akerblom I. E., Marron J. L., Mellon P. L. Mutual cross-interference between glucocorticoid receptor and CREB inhibits transactivation in placental cells. New Biol. 1992 May;4(5):527–540. [PubMed] [Google Scholar]
- 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]
- Vallejo M., Penchuk L., Habener J. F. Somatostatin gene upstream enhancer element activated by a protein complex consisting of CREB, Isl-1-like, and alpha-CBF-like transcription factors. J Biol Chem. 1992 Jun 25;267(18):12876–12884. [PubMed] [Google Scholar]
- Wagner S., Green M. R. HTLV-I Tax protein stimulation of DNA binding of bZIP proteins by enhancing dimerization. Science. 1993 Oct 15;262(5132):395–399. doi: 10.1126/science.8211160. [DOI] [PubMed] [Google Scholar]
- Yoshida M., Miyoshi I., Hinuma Y. Isolation and characterization of retrovirus from cell lines of human adult T-cell leukemia and its implication in the disease. Proc Natl Acad Sci U S A. 1982 Mar;79(6):2031–2035. doi: 10.1073/pnas.79.6.2031. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yoshida M., Osame M., Usuku K., Matsumoto M., Igata A. Viruses detected in HTLV-I-associated myelopathy and adult T-cell leukaemia are identical on DNA blotting. Lancet. 1987 May 9;1(8541):1085–1086. doi: 10.1016/s0140-6736(87)90506-x. [DOI] [PubMed] [Google Scholar]
- 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]