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Home / Equine Research Bank / J Virol / Mapping the sequences that mediate interaction of the equine...
Journal of virology2001; 75(21); 10219-10230; doi: 10.1128/JVI.75.21.10219-10230.2001

Mapping the sequences that mediate interaction of the equine herpesvirus 1 immediate-early protein and human TFIIB.

Abstract: The sole immediate-early (IE) gene of equine herpesvirus 1 encodes a 1,487-amino-acid (aa) regulatory phosphoprotein that independently activates expression of early viral genes. Coimmunoprecipitation assays demonstrated that the IE protein physically interacts with the general transcription factor TFIIB. Using a variety of protein-binding assays that employed a panel of IE truncation and deletion mutants expressed as in vitro-synthesized or glutathione S-transferase fusion proteins, we mapped a TFIIB-binding domain to aa 407 to 757 of the IE protein. IE mutants carrying internal deletions of aa 426 to 578 and 621 to 757 were partially defective for TFIIB binding, indicating that aa 407 to 757 may harbor more than one TFIIB-binding domain. The interaction between the IE protein and TFIIB is of physiological importance, as evidenced by transient-cotransfection assays. Partial deletion of the TFIIB-binding domain within the IE protein inhibited its ability to activate expression of the viral thymidine kinase gene, a representative early promoter, and of the IR5 gene, a representative late promoter, by greater than 20 and 50%, respectively. These results indicate that the interaction of the IE protein with TFIIB is necessary for its full transactivation function and that the IE-TFIIB interaction may be part of the mechanism by which the IE protein activates transcription.
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Publication Date: 2001-10-03 PubMed ID: 11581390PubMed Central: PMC114596DOI: 10.1128/JVI.75.21.10219-10230.2001Google 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 discusses the identification of the sequences in equine herpesvirus 1 immediate-early (IE) gene interacting with human TFIIB. Through various assays, the researchers established that the interaction is important for activating the expression of some early and late viral genes.

Understanding the Immediate-Early Protein and TFIIB Interaction

  • The study examines the IE gene of the equine herpesvirus 1, which encodes a complex regulatory phosphoprotein made up of 1,487 amino acids. This protein is crucial as it independently triggers the expression of early viral genes.
  • Researchers used coimmunoprecipitation assays to demonstrate that the IE protein physically interacts with a general transcription factor, TFIIB, in human cells. Essentially, they found that the IE protein binds with TFIIB, revealing a potentially important mechanism in the viral lifecycle.
  • The authors used a panel of IE truncation and deletion mutants for a range of protein-binding assays. This allowed them to map a TFIIB-binding domain in the IE protein’s sequence, located between amino acids 407 to 757.

Implications of the IE Protein and TFIIB Interaction

  • IE mutants with internal deletions of amino acids 426 to 578 and 621 to 757 were partially defective for TFIIB binding. This suggests that the span between amino acids 407 to 757 could contain more than one TFIIB-binding domain.
  • Using transient-cotransfection experiments, the researchers showed the physiological importance of the interaction between the IE protein and TFIIB.
  • They noted that a partial deletion of the TFIIB-binding domain within the IE protein hampered its ability to activate the expression of the viral thymidine kinase gene (an example of an early viral gene promoter) by more than 20%. The similar deletion also reduced activation of the IR5 gene (an example of a late viral gene promoter) by over 50%.
  • Conclusively, the research indicates the necessity of the IE protein’s interaction with TFIIB for its full transactivation function. The authors propose the IE-TFIIB interaction might form part of the mechanism through which the IE protein triggers transcription.

Cite This Article

APA
Jang HK, Albrecht RA, Buczynski KA, Kim SK, Derbigny WA, O'Callaghan DJ. (2001). Mapping the sequences that mediate interaction of the equine herpesvirus 1 immediate-early protein and human TFIIB. J Virol, 75(21), 10219-10230. https://doi.org/10.1128/JVI.75.21.10219-10230.2001

Publication

Journal of virology
ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 75
Issue: 21
Pages: 10219-10230

Researcher Affiliations

Jang, H K
  • Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130-3932, USA.
Albrecht, R A
    Buczynski, K A
      Kim, S K
        Derbigny, W A
          O'Callaghan, D J

            MeSH Terms

            • Animals
            • Binding Sites
            • Cells, Cultured
            • DNA / metabolism
            • Herpesvirus 1, Equid / chemistry
            • Herpesvirus 1, Equid / genetics
            • Humans
            • Immediate-Early Proteins / chemistry
            • Immediate-Early Proteins / metabolism
            • Mice
            • Precipitin Tests
            • Promoter Regions, Genetic
            • Transcription Factor TFIIB
            • Transcription Factors / metabolism
            • Transcriptional Activation

            Grant Funding

            • R01 AI022001 / NIAID NIH HHS
            • AI-22001 / NIAID NIH HHS

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