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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.
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Publication Date: 1993-04-01 PubMed ID: 8383228PubMed Central: PMC240285DOI: 10.1128/JVI.67.4.2064-2074.1993Google Scholar: Lookup
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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

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    This article has been cited 11 times.
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