FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / J Virol / Physical and functional characterization of transcriptional ...
Journal of virology1993; 67(4); 2064-2074; doi: 10.1128/JVI.67.4.2064-2074.1993

Physical and functional characterization of transcriptional control elements in the equine infectious anemia virus promoter.

Abstract: Equine infectious anemia virus (EIAV) is a lentivirus that causes a chronic disease of horses characterized by cyclic episodes of fever, anemia, and viremia. Although the genome and promoter of EIAV are much less complex than those of its relatives the primate immunodeficiency viruses, the cellular proteins that activate and regulate transcription of EIAV have not yet been identified. In this report, we show by electrophoretic mobility shift assays and DNase I footprinting that the EIAV promoter contains multiple binding sites for ubiquitous, cell type-specific, and inducible cellular proteins. Functional analysis by transient transfection of canine osteosarcoma (D17) and human epithelial carcinoma (HeLa) cells with EIAV promoters containing deletions or individually mutated DNA-binding sites demonstrated that these DNA-binding elements cooperatively regulate transcriptional activity. A methylated DNA-binding site (MDBP; also designated EF-C or EP) acts as either a positive or negative regulator of promoter activity, depending on the cell type or condition. Two PEA2 elements, an AP-1 site, and an ets/PEA3 motif confer a positive effect on promoter activity. The EIAV promoter is shown to be activated by treatment of HeLa cells with phorbol myristate acetate (PMA). DNA-binding activities were induced in PMA-treated HeLa cells and formed complexes on oligonucleotides that contain the EIAV AP-1 and ets/PEA3 elements. Functional analysis of mutated promoters indicated that the ets/PEA3 motif was the principal mediator of PMA activation.
Get Full Text
Publication Date: 1993-04-01 PubMed ID: 8383228PubMed Central: PMC240285DOI: 10.1128/JVI.67.4.2064-2074.1993Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses how the Equine infectious anemia virus (EIAV) promoter contains multiple binding sites for different cellular proteins, and how these DNA-binding elements cooperatively regulate transcriptional activity. It also studies the behaviour of this promoter under different conditions.

Overview of the Study

  • Equine Infectious Anemia Virus (EIAV) is a lentivirus causing chronic disease in horses involving episodes of fever, anemia, and viremia. This study is focussed on understanding the promoter of EIAV, specifically looking to identify the cellular proteins that help activate and regulate its transcription.
  • The researchers employed electrophoretic mobility shift assays and DNase I footprinting to show that the EIAV promoter consists of numerous binding sites for various cellular proteins, some ubiquitous, some cell type-specific, and others being inducible.

Functional Analysis

  • The team carried out functional analysis by transiently transfecting canine osteosarcoma (D17) and human epithelial carcinoma (HeLa) cells using EIAV promoters with either deletions or individually mutated DNA-binding sites.
  • The goal was to determine how these DNA-binding elements work together to regulate transcriptional activity. An interesting observation was the discovery of a methylated DNA-binding site that serves as either a positive or negative regulator of promoter activity, dependent on the cell type or condition.

Positive Effect on Promoter Activity

  • A unique discovery was that two PEA2 elements, an AP-1 site, and an ets/PEA3 motif can induce a positive effect on promoter activity.
  • Through further examination, it was also found that the EIAV promoter could be activated by treating HeLa cells with phorbol myristate acetate (PMA). This resulted in induction of DNA-binding activities in the treated HeLa cells which then formed complexes on oligonucleotides holding the EIAV AP-1 and ets/PEA3 elements.

Functional Analysis of Mutated Promoters

  • In the final stage of the study, mutated promoters were put through functional analysis. The researchers concluded that the ets/PEA3 motif played a central role in PMA activation.

Through this study, we gain valuable insights into the transcriptional control elements of the EIAV virus. This will potentially guide the development of more effective treatments for the viral infection in horses.

Cite This Article

APA
Carvalho M, Derse D. (1993). Physical and functional characterization of transcriptional control elements in the equine infectious anemia virus promoter. J Virol, 67(4), 2064-2074. https://doi.org/10.1128/JVI.67.4.2064-2074.1993

Publication

Journal of virology
ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 67
Issue: 4
Pages: 2064-2074

Researcher Affiliations

Carvalho, M
  • Laboratory of Viral Carcinogenesis, National Cancer Institute, Frederick Cancer Research and Development Center, Maryland 21702-1201.
Derse, D

    MeSH Terms

    • Animals
    • Base Sequence
    • Binding Sites
    • Cells, Cultured
    • DNA-Binding Proteins / metabolism
    • Dogs
    • Enhancer Elements, Genetic
    • Gene Expression Regulation, Viral
    • HeLa Cells
    • Horses
    • Humans
    • In Vitro Techniques
    • Infectious Anemia Virus, Equine / genetics
    • Methylation
    • Molecular Sequence Data
    • Oligodeoxyribonucleotides / chemistry
    • Promoter Regions, Genetic
    • Sequence Deletion
    • Tetradecanoylphorbol Acetate / pharmacology
    • Transcription Factors / metabolism
    • Transcription, Genetic

    References

    This article includes 38 references
    1. Takemoto KK, Furuno A, Kato K, Yoshiike K. Biological and biochemical studies of African green monkey lymphotropic papovavirus.. J Virol 1982 May;42(2):502-9.
      pubmed: 7086969doi: 10.1128/JVI.42.2.502-509.1982google scholar: lookup
    2. Hess JL, Small JA, Clements JE. Sequences in the visna virus long terminal repeat that control transcriptional activity and respond to viral trans-activation: involvement of AP-1 sites in basal activity and trans-activation.. J Virol 1989 Jul;63(7):3001-15.
      pubmed: 2542608doi: 10.1128/JVI.63.7.3001-3015.1989google scholar: lookup
    3. Kenney S, Natarajan V, Strike D, Khoury G, Salzman NP. JC virus enhancer-promoter active in human brain cells.. Science 1984 Dec 14;226(4680):1337-9.
      pubmed: 6095453doi: 10.1126/science.6095453google scholar: lookup
    4. Cheevers WP, McGuire TC. Equine infectious anemia virus: immunopathogenesis and persistence.. Rev Infect Dis 1985 Jan-Feb;7(1):83-8.
      pubmed: 2984759doi: 10.1093/clinids/7.1.83google scholar: lookup
    5. Veldman GM, Lupton S, Kamen R. Polyomavirus enhancer contains multiple redundant sequence elements that activate both DNA replication and gene expression.. Mol Cell Biol 1985 Apr;5(4):649-58.
      pubmed: 2985964doi: 10.1128/mcb.5.4.649-658.1985google scholar: lookup
    6. Kudo A, Ishihara T, Nishimura Y, Watanabe T. A cloned human immunoglobulin heavy chain gene with a novel direct-repeat sequence in 5' flanking region.. Gene 1985;33(2):181-9.
      pubmed: 3922855doi: 10.1016/0378-1119(85)90092-7google scholar: lookup
    7. Ostapchuk P, Diffley JF, Bruder JT, Stillman B, Levine AJ, Hearing P. Interaction of a nuclear factor with the polyomavirus enhancer region.. Proc Natl Acad Sci U S A 1986 Nov;83(22):8550-4.
      pubmed: 3022296doi: 10.1073/pnas.83.22.8550google scholar: lookup
    8. Derse D, Dorn PL, Levy L, Stephens RM, Rice NR, Casey JW. Characterization of equine infectious anemia virus long terminal repeat.. J Virol 1987 Mar;61(3):743-7.
      pubmed: 3027401doi: 10.1128/JVI.61.3.743-747.1987google scholar: lookup
    9. Kryszke MH, Piette J, Yaniv M. Induction of a factor that binds to the polyoma virus A enhancer on differentiation of embryonal carcinoma cells.. Nature 1987 Jul 16-22;328(6127):254-6.
      pubmed: 3037385doi: 10.1038/328254a0google scholar: lookup
    10. Piette J, Yaniv M. Two different factors bind to the alpha-domain of the polyoma virus enhancer, one of which also interacts with the SV40 and c-fos enhancers.. EMBO J 1987 May;6(5):1331-7.
      pubmed: 3038517doi: 10.1002/j.1460-2075.1987.tb02372.xgoogle scholar: lookup
    11. Garcia JA, Wu FK, Mitsuyasu R, Gaynor RB. Interactions of cellular proteins involved in the transcriptional regulation of the human immunodeficiency virus.. EMBO J 1987 Dec 1;6(12):3761-70.
      pubmed: 3428273doi: 10.1002/j.1460-2075.1987.tb02711.xgoogle scholar: lookup
    12. Dorn PL, Derse D. cis- and trans-acting regulation of gene expression of equine infectious anemia virus.. J Virol 1988 Sep;62(9):3522-6.
      pubmed: 2841502doi: 10.1128/JVI.62.9.3522-3526.1988google scholar: lookup
    13. Supakar PC, Weist D, Zhang DL, Inamdar N, Zhang XY, Khan R, Ehrlich KC, Ehrlich M. Methylated DNA-binding protein is present in various mammalian cell types.. Nucleic Acids Res 1988 Aug 25;16(16):8029-44.
      pubmed: 2901711doi: 10.1093/nar/16.16.8029google scholar: lookup
    14. Halazonetis TD, Georgopoulos K, Greenberg ME, Leder P. c-Jun dimerizes with itself and with c-Fos, forming complexes of different DNA binding affinities.. Cell 1988 Dec 2;55(5):917-24.
      pubmed: 3142692doi: 10.1016/0092-8674(88)90147-xgoogle scholar: lookup
    15. Wasylyk C, Imler JL, Wasylyk B. Transforming but not immortalizing oncogenes activate the transcription factor PEA1.. EMBO J 1988 Aug;7(8):2475-83.
      pubmed: 3142763doi: 10.1002/j.1460-2075.1988.tb03094.xgoogle scholar: lookup
    16. Fisch TM, Prywes R, Roeder RG. An AP1-binding site in the c-fos gene can mediate induction by epidermal growth factor and 12-O-tetradecanoyl phorbol-13-acetate.. Mol Cell Biol 1989 Mar;9(3):1327-31.
      pubmed: 2498646doi: 10.1128/mcb.9.3.1327-1331.1989google scholar: lookup
    17. Ben-Levy R, Faktor O, Berger I, Shaul Y. Cellular factors that interact with the hepatitis B virus enhancer.. Mol Cell Biol 1989 Apr;9(4):1804-9.
      pubmed: 2725524doi: 10.1128/mcb.9.4.1804-1809.1989google scholar: lookup
    18. Schönthal A, Büscher M, Angel P, Rahmsdorf HJ, Ponta H, Hattori K, Chiu R, Karin M, Herrlich P. The Fos and Jun/AP-1 proteins are involved in the downregulation of Fos transcription.. Oncogene 1989 May;4(5):629-36.
      pubmed: 2498806
    19. Duh EJ, Maury WJ, Folks TM, Fauci AS, Rabson AB. Tumor necrosis factor alpha activates human immunodeficiency virus type 1 through induction of nuclear factor binding to the NF-kappa B sites in the long terminal repeat.. Proc Natl Acad Sci U S A 1989 Aug;86(15):5974-8.
      pubmed: 2762307doi: 10.1073/pnas.86.15.5974google scholar: lookup
    20. Ostapchuk P, Scheirle G, Hearing P. Binding of nuclear factor EF-C to a functional domain of the hepatitis B virus enhancer region.. Mol Cell Biol 1989 Jul;9(7):2787-97.
      pubmed: 2550788doi: 10.1128/mcb.9.7.2787-2797.1989google scholar: lookup
    21. Timmers HT, Pronk GJ, Bos JL, van der Eb AJ. Analysis of the rat JE gene promoter identifies an AP-1 binding site essential for basal expression but not for TPA induction.. Nucleic Acids Res 1990 Jan 11;18(1):23-34.
      pubmed: 2106664doi: 10.1093/nar/18.1.23google scholar: lookup
    22. Dorn P, DaSilva L, Martarano L, Derse D. Equine infectious anemia virus tat: insights into the structure, function, and evolution of lentivirus trans-activator proteins.. J Virol 1990 Apr;64(4):1616-24.
      pubmed: 2157047doi: 10.1128/JVI.64.4.1616-1624.1990google scholar: lookup
    23. Boulukos KE, Pognonec P, Sariban E, Bailly M, Lagrou C, Ghysdael J. Rapid and transient expression of Ets2 in mature macrophages following stimulation with cMGF, LPS, and PKC activators.. Genes Dev 1990 Mar;4(3):401-9.
      pubmed: 2186967doi: 10.1101/gad.4.3.401google scholar: lookup
    24. Bhat NK, Thompson CB, Lindsten T, June CH, Fujiwara S, Koizumi S, Fisher RJ, Papas TS. Reciprocal expression of human ETS1 and ETS2 genes during T-cell activation: regulatory role for the protooncogene ETS1.. Proc Natl Acad Sci U S A 1990 May;87(10):3723-7.
      pubmed: 2187191doi: 10.1073/pnas.87.10.3723google scholar: lookup
    25. Dikstein R, Faktor O, Ben-Levy R, Shaul Y. Functional organization of the hepatitis B virus enhancer.. Mol Cell Biol 1990 Jul;10(7):3683-9.
      pubmed: 2355919doi: 10.1128/mcb.10.7.3683-3689.1990google scholar: lookup
    26. Gutman A, Wasylyk B. The collagenase gene promoter contains a TPA and oncogene-responsive unit encompassing the PEA3 and AP-1 binding sites.. EMBO J 1990 Jul;9(7):2241-6.
      pubmed: 2162765doi: 10.1002/j.1460-2075.1990.tb07394.xgoogle scholar: lookup
    27. van Dam H, Offringa R, Meijer I, Stein B, Smits AM, Herrlich P, Bos JL, van der Eb AJ. Differential effects of the adenovirus E1A oncogene on members of the AP-1 transcription factor family.. Mol Cell Biol 1990 Nov;10(11):5857-64.
      pubmed: 2172787doi: 10.1128/mcb.10.11.5857-5864.1990google scholar: lookup
    28. Zhang XY, Asiedu CK, Supakar PC, Khan R, Ehrlich KC, Ehrlich M. Binding sites in mammalian genes and viral gene regulatory regions recognized by methylated DNA-binding protein.. Nucleic Acids Res 1990 Nov 11;18(21):6253-60.
      pubmed: 2173824doi: 10.1093/nar/18.21.6253google scholar: lookup
    29. Carpenter S, Alexandersen S, Long MJ, Perryman S, Chesebro B. Identification of a hypervariable region in the long terminal repeat of equine infectious anemia virus.. J Virol 1991 Mar;65(3):1605-10.
      pubmed: 1847479doi: 10.1128/JVI.65.3.1605-1610.1991google scholar: lookup
    30. Ryseck RP, Bravo R. c-JUN, JUN B, and JUN D differ in their binding affinities to AP-1 and CRE consensus sequences: effect of FOS proteins.. Oncogene 1991 Apr;6(4):533-42.
      pubmed: 1827665
    31. Carvalho M, Derse D. Mutational analysis of the equine infectious anemia virus Tat-responsive element.. J Virol 1991 Jul;65(7):3468-74.
      pubmed: 1645778doi: 10.1128/JVI.65.7.3468-3474.1991google scholar: lookup
    32. Yoo W, Martin ME, Folk WR. PEA1 and PEA3 enhancer elements are primary components of the polyomavirus late transcription initiator element.. J Virol 1991 Oct;65(10):5391-400.
      pubmed: 1654447doi: 10.1128/JVI.65.10.5391-5400.1991google scholar: lookup
    33. Xin JH, Cowie A, Lachance P, Hassell JA. Molecular cloning and characterization of PEA3, a new member of the Ets oncogene family that is differentially expressed in mouse embryonic cells.. Genes Dev 1992 Mar;6(3):481-96.
      pubmed: 1547944doi: 10.1101/gad.6.3.481google scholar: lookup
    34. Dikstein R, Heffetz D, Ben-Neriah Y, Shaul Y. c-abl has a sequence-specific enhancer binding activity.. Cell 1992 May 29;69(5):751-7.
      pubmed: 1591775doi: 10.1016/0092-8674(92)90287-mgoogle scholar: lookup
    35. Seth A, Ascione R, Fisher RJ, Mavrothalassitis GJ, Bhat NK, Papas TS. The ets gene family.. Cell Growth Differ 1992 May;3(5):327-34.
      pubmed: 1633115
    36. Clark L, Hay RT. Sequence requirement for specific interaction of an enhancer binding protein (EBP1) with DNA.. Nucleic Acids Res 1989 Jan 25;17(2):499-516.
      pubmed: 2536920doi: 10.1093/nar/17.2.499google scholar: lookup
    37. Zhang XY, Supakar PC, Khan R, Ehrlich KC, Ehrlich M. Related sites in human and herpesvirus DNA recognized by methylated DNA-binding protein from human placenta.. Nucleic Acids Res 1989 Feb 25;17(4):1459-74.
      pubmed: 2537959doi: 10.1093/nar/17.4.1459google scholar: lookup
    38. Dignam JD, Lebovitz RM, Roeder RG. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.. Nucleic Acids Res 1983 Mar 11;11(5):1475-89.
      pubmed: 6828386doi: 10.1093/nar/11.5.1475google scholar: lookup

    Citations

    This article has been cited 11 times.
    1. de Pablo-Maiso L, Doménech A, Echeverría I, Gómez-Arrebola C, de Andrés D, Rosati S, Gómez-Lucia E, Reina R. Prospects in Innate Immune Responses as Potential Control Strategies against Non-Primate Lentiviruses.. Viruses 2018 Aug 17;10(8).
      doi: 10.3390/v10080435pubmed: 30126090google scholar: lookup
    2. Wang XF, Liu Q, Wang YH, Wang S, Chen J, Lin YZ, Ma J, Zhou JH, Wang X. Characterization of Equine Infectious Anemia Virus Long Terminal Repeat Quasispecies In Vitro and In Vivo.. J Virol 2018 Apr 15;92(8).
      doi: 10.1128/JVI.02150-17pubmed: 29386282google scholar: lookup
    3. Maury W, Thompson RJ, Jones Q, Bradley S, Denke T, Baccam P, Smazik M, Oaks JL. Evolution of the equine infectious anemia virus long terminal repeat during the alteration of cell tropism.. J Virol 2005 May;79(9):5653-64.
      doi: 10.1128/JVI.79.9.5653-5664.2005pubmed: 15827180google scholar: lookup
    4. Taube R, Fujinaga K, Irwin D, Wimmer J, Geyer M, Peterlin BM. Interactions between equine cyclin T1, Tat, and TAR are disrupted by a leucine-to-valine substitution found in human cyclin T1.. J Virol 2000 Jan;74(2):892-8.
      doi: 10.1128/jvi.74.2.892-898.2000pubmed: 10623752google scholar: lookup
    5. Cook RF, Leroux C, Cook SJ, Berger SL, Lichtenstein DL, Ghabrial NN, Montelaro RC, Issel CJ. Development and characterization of an in vivo pathogenic molecular clone of equine infectious anemia virus.. J Virol 1998 Feb;72(2):1383-93.
      doi: 10.1128/JVI.72.2.1383-1393.1998pubmed: 9445039google scholar: lookup
    6. Payne SL, Qi XM, Shao H, Dwyer A, Fuller FJ. Disease induction by virus derived from molecular clones of equine infectious anemia virus.. J Virol 1998 Jan;72(1):483-7.
      doi: 10.1128/JVI.72.1.483-487.1998pubmed: 9420249google scholar: lookup
    7. Maury W, Perryman S, Oaks JL, Seid BK, Crawford T, McGuire T, Carpenter S. Localized sequence heterogeneity in the long terminal repeats of in vivo isolates of equine infectious anemia virus.. J Virol 1997 Jul;71(7):4929-37.
      doi: 10.1128/JVI.71.7.4929-4937.1997pubmed: 9188555google scholar: lookup
    8. Carvalho M, Kirkland M, Derse D. Protein interactions with DNA elements in variant equine infectious anemia virus enhancers and their impact on transcriptional activity.. J Virol 1993 Nov;67(11):6586-95.
      doi: 10.1128/JVI.67.11.6586-6595.1993pubmed: 8411361google scholar: lookup
    9. Carvalho M, Derse D. The PU.1/Spi-1 proto-oncogene is a transcriptional regulator of a lentivirus promoter.. J Virol 1993 Jul;67(7):3885-90.
      doi: 10.1128/JVI.67.7.3885-3890.1993pubmed: 8389910google scholar: lookup
    10. Maury W. Monocyte maturation controls expression of equine infectious anemia virus.. J Virol 1994 Oct;68(10):6270-9.
      doi: 10.1128/JVI.68.10.6270-6279.1994pubmed: 8083967google scholar: lookup
    11. Sellon DC, Fuller FJ, McGuire TC. The immunopathogenesis of equine infectious anemia virus.. Virus Res 1994 May;32(2):111-38.
      doi: 10.1016/0168-1702(94)90038-8pubmed: 8067050google scholar: lookup
    Subscribe to our newsletter
    Join 99,359 horse owners like you who receive our email updates on horse health and nutrition.