title: Direct interaction of TFIIB and the IE protein of equine herpesvirus 1 is required for maximal trans-activation function.
abstract: Recently, we reported that the immediate-early (IE) protein of equine herpesvirus 1 (EHV-1) associates with transcription factor TFIIB [J. Virol. 75 (2001), 10219]. In the current study, the IE protein purified as a glutathione-S-transferase (GST) fusion protein was shown to interact directly with purified TFIIB in GST-pulldown assays. A panel of TFIIB mutants employed in protein-binding assays revealed that residues 125 to 174 within the first direct repeat of TFIIB mediate its interaction with the IE protein. This interaction is physiologically relevant as transient transfection assays demonstrated that (1). exogenous native TFIIB did not perturb IE protein function, and (2). ectopic expression of a TFIIB mutant that lacked the IE protein interactive domain significantly diminished the ability of the IE protein to trans-activate EHV-1 promoters. These results suggest that an interaction of the IE protein with TFIIB is an important aspect of the regulatory role of the IE protein in the trans-activation of EHV-1 promoters.
authors.affiliation: Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932, USA.
authors.forename: Randy A
authors.initials: RA
authors.lastname: Albrecht
authors.forename: Hyung K
authors.initials: HK
authors.lastname: Jang
authors.forename: Seong K
authors.initials: SK
authors.lastname: Kim
authors.forename: Dennis J
authors.initials: DJ
authors.lastname: O'Callaghan
mesh_terms: D000818:Animals; D001665:Binding Sites; D002478:Cells, Cultured; D017781:Genes, Immediate-Early; D004861:Herpesvirus 1, Equid / Q000235:genetics / Q000502:physiology*; D017874:Immediate-Early Proteins / Q000737:chemistry / Q000502:physiology*; D051379:Mice; D035581:Transcription Factor TFIIB / Q000502:physiology*; D015533:Transcriptional Activation
chemical_list: D017874:Immediate-Early Proteins; D035581:Transcription Factor TFIIB