Skip to main content
SpringerLink
Log in

NF-κB site-mediated negative regulation of the HIV-1 promoter by CCAAT/enhancer binding proteins in brain-derived cells

  • Original Article
  • Published:
Journal of Molecular Neuroscience Aims and scope Submit manuscript

Abstract

Several transcription regulatory elements that interact with cellular DNA-binding proteins have been identified in the HIV-1 long terminal repeat (LTR). We have identified two sequence motifs in the U3 region of the LTR that are similar to the consensus 9-bp DNA-binding element of the CCAAT/enhancer-binding protein (C/EBP) family of transcription factors. One of the sequences (promoter-proximal) mapped immediately upstream of the NF-κB element, whereas the other (promoter-distal) completely overlapped the upstream stimulatory factor (USF) binding site. In this study, we investigated the role of the enhancer-proximal consensus C/EBP binding sequence in the expression of the HIV-1 LTR. In cotransfection assays we found that although this sequence is a functional C/EBP-responsive element, the regulation of the HIV promoter by C/EBP is very complex. C/EBP isoforms inhibited the phorbol 12-myristate 13-acetate (PMA)-stimulated HIV-1 promoter activity in human glioblastoma U138MG and neuroblastoma SHSY5Y cells, but not in HeLa epithelial cells, and this inhibition required the NF-κB element. C/EBP also downregulated the HIV NF-κB element-containing SV40 early promoter activity, regardless of the presence of the flanking C/EBP-binding sequences, in the two brain-derived cells. In electrophoretic mobility shift assays with nuclear extracts from HeLa and U138MG cells, purified C/EBP markedly increased the complex formation between endogenous proteins and the NF-κB DNA probe without detectable association with the complex. However, with extracts from U138MG cells but not from HeLa cells, a slow migrating complex was observed. Our data suggest that the C/EBP family of transcription factors can downregulate the HIV-1 promoter activity in CNS-derived cells through the NF-κB binding elements.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Akira S., Isshiki H., Sugita T., Tanabe O., Kinoshita S., Nishio Y., Nakajima T., Hirano T., and Kishimoto T. (1990) A nuclear factor for IL-6 expression (NF-IL-6) is a member of a C/EBP family.EMBO J. 9, 1897–1906.

    PubMed  CAS  Google Scholar 

  • Atwood W. J., Berger J. R., Kaderman R., Tornatore C. S., and Major E. O. (1993) Human immunodeficiency virus type 1 infection of the brain.Clin. Microbiol. Rev. 6, 339–366.

    PubMed  CAS  Google Scholar 

  • Baeuerle P. A. and Baltimore D. (1989) A 65 kD subunit of active NF-kB is required for inhibition of NF-kB by IkB.Genes Dev. 3, 1689–1698.

    Article  PubMed  CAS  Google Scholar 

  • Bagasra O., Khalili K., Seshamma T., Taylor J. P., and Pomerantz R. J. (1992) TAR-independent replication of human immunodeficiency virus type 1 in glial cells.J. Viorl. 66, 7522–7528.

    CAS  Google Scholar 

  • Berkhout B. and Jeang K.-T. (1992) Functional roles for the TATA promoter and enhancers in basal and Tat-induced expression of the human immunodeficiency virus type 1 long terminal repeat.J. Virol. 66, 139–149.

    PubMed  CAS  Google Scholar 

  • Birkenmeier E. H., Gwynn B., Howard S., Jerry J., Gordon J. I., Landschulz W. H., and McKnight S. L. (1989) Tissue-specific expression, developmental regulation, and genetic mapping of the gene encoding CCAAT/enhancer binding protein.Genes Dev. 3, 1146–1156.

    Article  PubMed  CAS  Google Scholar 

  • Breen E. C., Rezai A. R., Nakajima K., Beall G. N., Mitsuyasu R. T., Hirano T., Kishimoto T., and Martinez-Maza O. (1990) Infection of HIV is associated with elevated IL-6 levels and production.J. Immunol. 144, 480–484.

    PubMed  CAS  Google Scholar 

  • Cao Z., Umek R. M., and McKnight S. L. (1991) Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-LI cells.Genes Dev. 5, 1538–1552.

    Article  PubMed  CAS  Google Scholar 

  • Chang C.-J., Chen T.-T., and Lei H.-Y., Chen D. S., and Lee S.-C. (1990) Molecular cloning of a transcription factor, AGP/EBP, that belongs to members of the C/EBP family.Mol. Cell. Biol. 10, 6642–6653.

    PubMed  CAS  Google Scholar 

  • Christy R. J., Yang V. W., Ntambi J. M., Geiman D. E., Landschulz W. H., Friedman A. D., Nakabeppu Y., Kelly T. J., and Lane M. D. (1989) Differentiation-induced gene expression in 3T3-L1 preadipocytes: CCAAT/enhancer binding protein interacts with and activates the promoters of two adipocyte-specific genes.Genes Dev. 3, 1323–1335.

    Article  PubMed  CAS  Google Scholar 

  • Descombes P., Chojkier M., Lichsteiner S., Falvey E., and Schibler U. (1990) LAP, a novel member of the C/EBP gene family, encodes a liver-enriched transcriptional activator protein.Genes Dev. 4, 1541–1551.

    Article  PubMed  CAS  Google Scholar 

  • Dignam J. D., Lebovitz R. M., and Roeder R. G. (1983) Accurate transcription initiation by RNA polymerase 11, in a soluble extract from isolated mammalian nuclei.Nucleic Acids Res. 11, 1475–1489.

    Article  PubMed  CAS  Google Scholar 

  • Friedman A. D., Landschulz W. H., and McKnight S. L. (1989) CCAAT/enhancer binding protein activates the promoter of the serum albumin gene in cultured hepatoma cells.Genes Dev. 3, 1314–1322.

    Article  PubMed  CAS  Google Scholar 

  • Fujita T., Nolan G. P., Ghosh S., and Baltimore D. (1992) Independent modes of transcriptional activation by the p50 and p65 subunits of NF-kB.Genes Dev. 6, 775–787.

    Article  PubMed  CAS  Google Scholar 

  • Gallo P., Frei K., Rordorf C., Lazdins J., Tavolato B., and Fontana A. (1989) Human immunodeficiency virus type 1 (HIV-1) infection of the central nervous system: an evaluation of cytokines in cerebrospinal fluid.J. Neuroimmunol. 23, 109–116.

    Article  PubMed  CAS  Google Scholar 

  • Garcia J. A., Harrich D., Soultanakis E., Wu F., Mitsuyasu R., and Gaynor R. B. (1989) Human immunodeficiency virus type 1 LTR TATA and TAR region sequences required for transcriptional regulation.EMBO J. 8, 765–778.

    PubMed  CAS  Google Scholar 

  • Garcia J. A., Wu F. K., Mitsuyasu R., and Gaynor R. B. (1987) Interactions of cellular proteins involved in the transcriptional regulation of the human immunodeficiency virus.EMBO J. 6, 3761–3770.

    PubMed  CAS  Google Scholar 

  • Gartner S., Markovits P., Markovitz D. M., Kaplan M. H., Gallo R. C., and Popovic M. (1986) The role of mononuclear phagocytes in HTLV-III/LAV infection.Science 233, 215–219.

    Article  PubMed  CAS  Google Scholar 

  • Gaynor R. (1992) Cellular transcription factors involved in the regulation of HIV-1 gene expression.AIDS 6, 347–363.

    Article  PubMed  CAS  Google Scholar 

  • Gorman C. M., Merlino G. T., Willingham M. C., Pastan I., and Howard B. H. (1982) The rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection.Proc. Natl. Acad. Sci. USA 79, 6777–6781.

    Article  PubMed  CAS  Google Scholar 

  • Graves B. J., Johnson P. F., and McKnight S. L. (1986) Homologous recognition of a promoter domain common to the MSV LTR and the HSV tk gene.Cell 44, 565–576.

    Article  PubMed  CAS  Google Scholar 

  • Grove M. and Plumb M. (1993) C/EBP, NF-κB, and c-Ets family members and transcriptional regulation of the cell-specific and inducible macrophage inflammatory protein 1α immediate-early gene.Mol. Cell. Biol. 13, 5276–5289.

    PubMed  CAS  Google Scholar 

  • Harrich D., Garcia J., Wu F., Mitsuyasu R., Gonzalez J., and Gaynor R. (1989) Role of SPI-binding domains inin vivo transcriptional regulation of the human immunodeficiency virus type 1 long terminal repeat.J. Virol. 63, 2585–2591.

    PubMed  CAS  Google Scholar 

  • Herrera R., Ro H. S., Robinson G. S., Xanthopoulos K. G., and Spiegelman B. M. (1989) A direct role for C/EBP and the AP-1-binding site in gene expression linked to adipocyte differentiation.Mol. Cell. Biol. 9, 5331–5339.

    PubMed  CAS  Google Scholar 

  • Johnson P. F., Landschulz W. H., Graves B. J., and McKnight S. L. (1987) Identification of a rat liver nuclear protein that binds to the enhancer core element of three animal viruses.Genes Dev. 1, 133–146.

    Article  PubMed  CAS  Google Scholar 

  • Kaufman J. D., Valandra G., Roderiquez G., Bushar G., Giri C., and Norcross M. A. (1987) Phorbol ester enhances human immunodeficiency virus-promoted gene expression and acts on a repeated 10-base-pair functional enhancer element.Mol. Cell. Biol. 7, 3759–3766.

    PubMed  CAS  Google Scholar 

  • Kawakami K., Scheidereit C., and Roeder R. (1988) Identification and purification of a human immunoglobulin enhancer-binding protein (NFκB) that activates transcription from a human immunodeficiency virus type 1 promoter in vitro.Proc. Natl. Acad. Sci. USA 85, 4700–4704.

    Article  PubMed  CAS  Google Scholar 

  • Korner M., Rattner A., Mauxion F., Sen R., and Citri Y. (1989) A brain-specific transcription activator.Neuron 3, 563–572.

    Article  PubMed  CAS  Google Scholar 

  • Laimins L. A., Gruss P., Pozzati R., and Khoury G. (1984) Characterization of enhancer elements in the long terminal repeat of Moloney murine sarcoma virus.J. Virol. 49, 183–189.

    PubMed  CAS  Google Scholar 

  • Landschulz W. H., Johnson P. F., and McKnight S. L. (1988) The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins.Science 240, 1759–1764.

    Article  PubMed  CAS  Google Scholar 

  • LeClair K. P., Blanar M. A., and Sharp P. A. (1992) The p50 subunit of NF-κB associates with the NF-IL-6 transcription factor.Proc. Natl. Acad. Sci. USA 89, 8145–8149.

    Article  PubMed  CAS  Google Scholar 

  • Liberman T. A. and Baltimore D. (1990) Activation of interleukin-6 gene expression through the NF-κB transcription factor.Mol. Cell. Biol. 10, 2327–2334.

    Google Scholar 

  • Matsusaka T., Fujikawa K., Nishio Y., Mukaida N., Matsushima K., Kishimoto T., and Akira S. (1993) Transcriptional factors NF-IL-6 and NF-κB synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8.Proc. Natl. Acad. Sci. USA 90, 10,193–10,197.

    Article  CAS  Google Scholar 

  • McKnight S. L., Lane M. D., and Gluecksohn-Waelsh S. (1989) Is CCAAT/enhancer binding protein a central regulator of energy metabolism?Genes Dev. 3, 2021–2024.

    Article  PubMed  CAS  Google Scholar 

  • Mukaida N., Mahe Y., and Matsushima K. (1990) Cooperative interaction of nuclear factor-κB and cis-regulatory enhancer binding protein-like factor binding elements in activating the interleukin-8 gene by proinflammatory cytokines.J. Biol. Chem. 265, 21,128–21,133.

    CAS  Google Scholar 

  • Nabel G. and Baltimore D. (1987) An inducible transcription factor activates expression of human immunodeficiency virus in T-cells.Nature (London) 326, 711–713.

    Article  CAS  Google Scholar 

  • Nakajima K., Martinez-Maza O., Hirano T., Breen E. C., Nishanian P. G., Salazar-Gonzalez J. F., Fahey J. L., and Kishimoto T. (1989) Induction of IL-6 (B cell stimulatory factor-2: IFN-β2) production by HIV.J. Immunol. 142, 531–536.

    PubMed  CAS  Google Scholar 

  • Nicholson J. K., Cross G. D., Callaway C. S., and McDougal J. S. (1986)In vitro infection of human monocytes with human T lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV).J. Immunol. 137, 323–329.

    PubMed  CAS  Google Scholar 

  • Nickoloff B. J., Karabin G. D., Barker J. N. W. N., Griffiths C. E. M., Sarma V., Mitra R. S., Elder J. T., Kunkel S. L., and Dixit V. M. (1991) Cellular localization of interleukin-8 and its inducer, tumor necrosis factor-alpha in psoriasis.Am. J. Pathol. 138, 129–140.

    PubMed  CAS  Google Scholar 

  • Nordeen S. K., Green P. P. III, and Fowlkes D. M. (1987) A rapid, sensitive and inexpensive assay for chloramphenicol acetyltransferase.DNA 6, 173–178.

    Article  PubMed  CAS  Google Scholar 

  • Oliveira L. C., Mukaida N., Matsushima K., and Vilcek J. (1994) Transcriptional inhibition of the interleukin-8 gene by interferon is mediated by the NF-κB site.Mol. Cell. Biol. 14, 5300–5308.

    PubMed  CAS  Google Scholar 

  • Osborn L., Kunkel S., and Nabel G. J. (1989) Tumor necrosis factor α and interleukin 1 stimulate the human immunodeficiency virus enhancer by activation of the nuclear factor κB.Proc. Natl. Acad. Sci. USA 86, 2336–2340.

    Article  PubMed  CAS  Google Scholar 

  • Poli V., Mancini F. P., and Cortese R. (1990) IL-6DBP, a nuclear protein involved in interleukin-6 signal transduction, defines a new family of leucine zipper proteins related to C/EBP.Cell 63, 643–653.

    Article  PubMed  CAS  Google Scholar 

  • Rando R. F., Pellet P. E., Luciw P. A., Bohan C. A., and Srinivasan A. (1987) Transactivation of human immunodeficiency virus, by herpes viruses.Oncogene 1, 13–18.

    PubMed  CAS  Google Scholar 

  • Ratner L., Haseltine W., Patarca R., Livak K. J., Starcich B., Josephs S. F., Doran E. R., Rafalski A., Whitehorn E. A., Baumeister K., Ivanoff L., Petteway S. R. Jr., Pearson M. L., Lautenberger J. A., Papas T. S., Ghrayeb J., Chang N. T., Gallo R. C., and Wong-Staal F. (1985) Complete nucleotide sequence of the AIDS virus, HTLV-III.Nature (London) 313, 277–284.

    Article  CAS  Google Scholar 

  • Rattner A., Korner M., Walker M. D., and Citri Y. (1993) NF-κB activates the HIV promoter in neurons.EMBO J. 12, 4261–4267.

    PubMed  CAS  Google Scholar 

  • Roman C., Platero J. S., Shuman J., and Calame K. (1990) Ig/EBP-1: a ubiquitously expressed immunoglobulin enhancer binding protein that is similar to C/EBP and heterodimerizes with C/EBP.Genes Dev. 4, 1404–1415.

    Article  PubMed  CAS  Google Scholar 

  • Ryden T. A. and Beemon K. (1989) Avian retroviral long terminal repeats bind CCAAT/enhancer-binding protein.Mol. Cell. Biol. 9, 1155–1164.

    PubMed  CAS  Google Scholar 

  • Sanchez-Pescador R., Power M. D., Barr P. J., Steimer K. S., Stempien M. M., Brown-Shimer S. L., Gee W. W., Renard A., Randolph A., Levy J. A., Dina D., and Luciw P. A. (1985) Nucleotide sequence and expression of an AIDS-associated retrovirus (ARV-2).Science 227, 484–492.

    Article  PubMed  CAS  Google Scholar 

  • Scala G., Ruocco M. R., Ambrosino C., Mallardo M., Giordano V., Baldassarre F., Dragonetti E., Quinto I., and Ventura S. (1994) The expression of the interleukin 6 gene is induced by the human immunodeficiency virus 1 Tat protein.J. Exp. Med. 179, 961–971.

    Article  PubMed  CAS  Google Scholar 

  • Stein B. and Baldwin A. S. Jr. (1993) Distinct mechanisms for regulation of the interleukin-8 gene involve synergism and cooperativity between C/EBP and NF-κB.Mol. Cell. Biol. 13, 7191–7198.

    PubMed  CAS  Google Scholar 

  • Stein B., Cogswell P. C., and Baldwin A. S. Jr. (1993) Functional and physical associations between NF-κB and C/EBP family members: a rel domain-bZIP interaction.Mol. Cell Biol. 13, 3964–3974.

    PubMed  CAS  Google Scholar 

  • Taylor J. P., Pomerantz R., Bagasra O., Chawdhury M., Rappaport J., Khalili K., and Amini S. (1992) TAR-independent transactivation by Tat in cells derived from the CNS: a novel mechanism of HIV-1 gene regulation.EMBO J. 11, 3395–3403.

    PubMed  CAS  Google Scholar 

  • Taylor J. P., Pomerantz R. J., Raj G. V., Kashanchi F., Brady J. N., Amini S., and Khalili K. (1994) Central nervous system-derived cells express a κB binding activity that enhances human immunodeficiency virus type 1 transactivationin vitro and facilitates TAR-independent transactivation byTat.J. Virol. 68, 3971–3981.

    PubMed  CAS  Google Scholar 

  • Tesmer V. M., Rajadhyaksha A., Babin J., and Bina M. (1993) NF-IL-6-mediated transcriptional activation of the long terminal repeat of the human immunodeficiency virus type 1.Proc. Natl. Acad. Sci. USA 90, 7298–7302.

    Article  PubMed  CAS  Google Scholar 

  • Thomassin H., Hamel D., Bernier D., Guertin M., and Belanger L. (1992) Molecular cloning of two C/EBP-related proteins that bind to the promoter and the enhancer of the alfafetoprotein gene. Further analysis of the C/EBPβ and C/EBPγ.Nucleic Acids Res. 20, 3091–3098.

    Article  PubMed  CAS  Google Scholar 

  • Umek R. M., Friedman A. D., and McKnight S. L. (1991) CCAAT/enhancer binding protein: a component of a differentiation switch.Science 251, 288–292.

    Article  PubMed  CAS  Google Scholar 

  • Urban M. B., Schreck R., and Baeuerle A. (1991) NF-κB contacts DNA by a heterodimer of the p50 and p65 subunit.EMBO J. 10, 1817–1825.

    PubMed  CAS  Google Scholar 

  • Vincenti M. P., Burrell T. A., and Taffet S. M. (1992) Regulation of NF-kappa B activity in murine macrophages: effect of bacterial lipopolysaccharide and phorbol ester.J. Cell. Physiol. 150, 204–213.

    Article  PubMed  CAS  Google Scholar 

  • Vinson C. R., Sigler P. B., and McKnight S. L. (1989) Scissors-grip model for DNA recognition by a family of leucine zipper proteins.Science 246, 911–916.

    Article  PubMed  CAS  Google Scholar 

  • Wain-Hobson S., Sonigo P., Danos O., Cole S., and Alizon M. (1985) Nucleotide sequence of the AIDS virus LAV.Cell 40, 9–17.

    Article  PubMed  CAS  Google Scholar 

  • Williams S. C., Cantwell C. A., and Johnson P. F. (1991) A family of C/EBP-related proteins capable of forming covalently linked leucine zipper dimersin vitro.Genes Dev. 5, 1553–1567.

    Article  PubMed  CAS  Google Scholar 

  • Zeichner S. L., Kim J. Y. H., and Alwine J. C. (1991) Linker-scanning mutational analysis of the transcriptional activity of the human immunodeficiency virus type 1 long terminal repeat.J. Virol. 65, 2436–2444.

    PubMed  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Debasis Mondal

    Present address: Department of Pharmacology, Tulane University Medical Center, 70112, New Orleans, LA

Authors and Affiliations

  1. Departments of Molecular Oncology, Alton Ochsner Medical Foundation, 70121, New Orleans, LA

    Debasis Mondal & Om Prakash

  2. Departments of Molecular Genetics, Alton Ochsner Medical Foundation, 70121, New Orleans, LA

    Jawed Alam

  3. Departments of Microbiology, Immunology and Parasitology, LSU Medical Center, 70119, New Orleans, LA

    Debasis Mondal & Om Prakash

  4. Departments of Biochemistry and Molecular Biology, LSU Medical Center, 70119, New Orieans, LA

    Jawed Alam & Om Prakash

Authors
  1. Debasis Mondal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jawed Alam
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Om Prakash
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

{ogDepartment of Pharmacology} {onTualen University Medical Center} {cxNew Orleans} {czLA} {pc70112}

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mondal, D., Alam, J. & Prakash, O. NF-κB site-mediated negative regulation of the HIV-1 promoter by CCAAT/enhancer binding proteins in brain-derived cells. J Mol Neurosci 5, 241–258 (1994). https://doi.org/10.1007/BF02736725

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02736725

Index Entries

Search

Navigation