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Home / Equine Research Bank / J Virol / Delineating minimal protein domains and promoter elements fo...
Journal of virology1995; 69(4); 2605-2610; doi: 10.1128/JVI.69.4.2605-2610.1995

Delineating minimal protein domains and promoter elements for transcriptional activation by lentivirus Tat proteins.

Abstract: Lentivirus Tat proteins comprise a novel class of RNA-binding transcriptional activators that are essential for viral replication. In this study, we performed a series of protein fusion experiments to delineate the minimal protein domains and promoter elements required for Tat action. We show that a 15-amino-acid region of equine infectious anemia virus (EIAV) Tat protein, when fused to the GAL4 or LexA DNA binding domain, can activate transcription in appropriate promoter contexts. In the natural human immunodeficiency virus type 1 long terminal repeat, activation by Tat is dependent on multiple binding sites for the cellular transcription factor SP1. We delineate a 114-amino-acid region of the SP1 glutamine-rich activation domain that when fused to the GAL4 DNA binding domain can support transcription activation by Tat. Using these Tat and SP1 derivatives, we show that Tat activation can be reconstructed on a completely synthetic promoter lacking all cis-acting elements unique to the human immunodeficiency virus long terminal repeat. Our results indicate that lentivirus Tat proteins have essential properties of typical cellular transcriptional activators and define useful reagents for studying the detailed mechanism of Tat action.
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Publication Date: 1995-04-01 PubMed ID: 7884911PubMed Central: PMC188939DOI: 10.1128/JVI.69.4.2605-2610.1995Google Scholar: Lookup
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  • Non-U.S. Gov't
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  • U.S. Gov't
  • P.H.S.

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 investigates the necessary protein domains and promoter elements for transcriptional activation by lentivirus Tat proteins, which are essential for viral replication. Through experiments, the researchers were able to define important constructs for further study on how Tat proteins function.

Protein Fusion Experiments

  • The research involves executing a series of protein fusion experiments. This is done to identify the minimal protein domains and promoter elements necessary for the function of Tat, a lentivirus protein that plays a vital role in viral replication.
  • One of the significant findings was that a minor 15-amino-acid region of equine infectious anemia virus (EIAV) Tat protein, when combined with the GAL4 or LexA DNA binding domain, had the capacity to initiate transcription in appropriate promoter settings.

Transcription Activation Dependency

  • In natural human immunodeficiency virus (HIV-1) long terminal repeat, activation is dependent on multiple binding sites for a cellular transcription factor known as SP1.
  • The researchers were able to decipher a region of the SP1 activation domain that, when combined with the GAL4 DNA binding domain, can support transcription activation by Tat. This SP1 activation domain was glutamine-rich and made up of 114 amino acids.

Reconstructed Tat Activation

  • The article explains how Tat activation was restructured on a completely artificial promoter that lacked all cis-acting elements found in the human immunodeficiency virus long terminal repeat. This was achieved by using the defined Tat and SP1 derivatives discovered in previous experiments.
  • Therefore, lentivirus Tat proteins possess essential characteristics of typical cellular transcriptional activators. This realization is crucial to creating useful reagents to investigate the detailed mechanism of Tat action.

Cite This Article

APA
Southgate CD, Green MR. (1995). Delineating minimal protein domains and promoter elements for transcriptional activation by lentivirus Tat proteins. J Virol, 69(4), 2605-2610. https://doi.org/10.1128/JVI.69.4.2605-2610.1995

Publication

Journal of virology
ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 69
Issue: 4
Pages: 2605-2610

Researcher Affiliations

Southgate, C D
  • Howard Hughes Medical Institute, Program in Molecular Medicine, University of Massachusetts Medical Center 01605.
Green, M R

    MeSH Terms

    • Animals
    • Cricetinae
    • DNA-Binding Proteins
    • Fungal Proteins / metabolism
    • Gene Products, tat / metabolism
    • HIV Long Terminal Repeat
    • HIV-1 / metabolism
    • HeLa Cells
    • Humans
    • Infectious Anemia Virus, Equine / genetics
    • Infectious Anemia Virus, Equine / metabolism
    • Lentivirus / genetics
    • Lentivirus / metabolism
    • Promoter Regions, Genetic
    • Saccharomyces cerevisiae Proteins
    • Transcription Factors / metabolism
    • Transcriptional Activation
    • tat Gene Products, Human Immunodeficiency Virus

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    Citations

    This article has been cited 10 times.
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