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

H. K. Jang, R. A. Albrecht, K. A. Buczynski, S. K. Kim, Wilbert Derbigny, D. J. O'Callaghan

Research output: Contribution to journalArticle

23 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)10219-10230
Number of pages12
JournalJournal of Virology
Volume75
Issue number21
DOIs
StatePublished - 2001
Externally publishedYes

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Transcription Factor TFIIB
Equid Herpesvirus 1
Immediate-Early Proteins
Equid herpesvirus 1
proteins
amino acids
Amino Acids
assays
genes
promoter regions
thymidine kinase
General Transcription Factors
mutants
phosphoproteins
protein binding
transcriptional activation
Immediate-Early Genes
Viral Genes
Thymidine Kinase
Phosphoproteins

ASJC Scopus subject areas

  • Immunology

Cite this

Jang, H. K., Albrecht, R. A., Buczynski, K. A., Kim, S. K., Derbigny, W., & O'Callaghan, D. J. (2001). Mapping the sequences that mediate interaction of the equine herpesvirus 1 immediate-early protein and human TFIIB. Journal of Virology, 75(21), 10219-10230. https://doi.org/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. / Jang, H. K.; Albrecht, R. A.; Buczynski, K. A.; Kim, S. K.; Derbigny, Wilbert; O'Callaghan, D. J.

In: Journal of Virology, Vol. 75, No. 21, 2001, p. 10219-10230.

Research output: Contribution to journalArticle

Jang, HK, Albrecht, RA, Buczynski, KA, Kim, SK, Derbigny, W & O'Callaghan, DJ 2001, 'Mapping the sequences that mediate interaction of the equine herpesvirus 1 immediate-early protein and human TFIIB', Journal of Virology, vol. 75, no. 21, pp. 10219-10230. https://doi.org/10.1128/JVI.75.21.10219-10230.2001
Jang, H. K. ; Albrecht, R. A. ; Buczynski, K. A. ; Kim, S. K. ; Derbigny, Wilbert ; O'Callaghan, D. J. / Mapping the sequences that mediate interaction of the equine herpesvirus 1 immediate-early protein and human TFIIB. In: Journal of Virology. 2001 ; Vol. 75, No. 21. pp. 10219-10230.
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