Structural and antigenic identification of the ORF12 protein (αTIF) of equine herpesvirus 1

Jill B. Lewis, Yeini G. Thompson, Xuan Feng, V. Roger Holden, Dennis O'Callaghan, Gretchen B Caughman

Research output: Contribution to journalArticle

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Abstract

The equine herpesvirus 1 (EHV-1) homolog of the herpes simplex virus type 1 (HSV-1) tegument phosphoprotein, αTIF (Vmw65; VP16), was identified previously as the product of open reading frame 12 (ORF12) and shown to transactivate immediate early (IE) gene promoters. However, a specific virion protein corresponding to the ORF12 product has not been identified definitively. In the present study the ORF12 protein, designated ETIF, was identified as a 60-kDa virion component on the basis of protein fingerprint analyses in which the limited proteolysis profiles of the major 60-kDa in vitro transcription/translation product of an ORF12 expression vector (pT7-12) were compared to those of purified virion proteins of similar size. ETIF was localized to the viral tegument in Western blot assays of EHV-1 virions and subvirion fractions using polyclonal antiserum and monoclonal antibodies generated against a glutathione-S-transferase-ETIF fusion protein. Northern and Western blot analyses of EHV-1-infected cell lysates prepared under various metabolic blocks indicated that ORF12 is expressed as a late gene, and cross reaction of polyclonal anti-GST-ETIF with a 63.5-kDa HSV-1 protein species suggested that ETIF and HSV-1 αTIF are antigenically related. Last, DNA band shift assays used to assess ETIF-specific complex formation indicated that ETIF participates in an infected cell protein complex with the EHV-1 IE promoter TAATGARAT motif.

Original languageEnglish (US)
Pages (from-to)369-375
Number of pages7
JournalVirology
Volume230
Issue number2
DOIs
StatePublished - Apr 14 1997

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Equid Herpesvirus 1
Open Reading Frames
Virion
Human Herpesvirus 1
Proteins
Western Blotting
Immediate-Early Genes
Peptide Mapping
Phosphoproteins
Cross Reactions
Glutathione Transferase
Northern Blotting
Proteolysis
Immune Sera
Monoclonal Antibodies
DNA
Genes

ASJC Scopus subject areas

  • Virology

Cite this

Structural and antigenic identification of the ORF12 protein (αTIF) of equine herpesvirus 1. / Lewis, Jill B.; Thompson, Yeini G.; Feng, Xuan; Holden, V. Roger; O'Callaghan, Dennis; Caughman, Gretchen B.

In: Virology, Vol. 230, No. 2, 14.04.1997, p. 369-375.

Research output: Contribution to journalArticle

Lewis, JB, Thompson, YG, Feng, X, Holden, VR, O'Callaghan, D & Caughman, GB 1997, 'Structural and antigenic identification of the ORF12 protein (αTIF) of equine herpesvirus 1', Virology, vol. 230, no. 2, pp. 369-375. https://doi.org/10.1006/viro.1997.8477
Lewis, Jill B. ; Thompson, Yeini G. ; Feng, Xuan ; Holden, V. Roger ; O'Callaghan, Dennis ; Caughman, Gretchen B. / Structural and antigenic identification of the ORF12 protein (αTIF) of equine herpesvirus 1. In: Virology. 1997 ; Vol. 230, No. 2. pp. 369-375.
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