title: Requirement of the N-terminal activation domain of herpes simplex virus ICP4 for viral gene expression.
abstract: ICP4 is the major activator of herpes simplex virus (HSV) transcription. Previous studies have defined several regions of ICP4 that are important for viral gene expression, including a DNA binding domain and transactivation domains that are contained in the C-terminal and N-terminal 520 and 274 amino acids, respectively. Here we show that the N-terminal 210 amino acids of ICP4 are required for interactions with components of TFIID and mediator and, as a consequence, are necessary for the activation of viral genes. A mutant of ICP4 deleted for amino acids 30 to 210, d3-10, was unable to complement an ICP4 null virus at the level of viral replication. This was the result of a severe deficiency in viral gene and protein expression. The absence of viral gene expression coincided with a defect in the recruitment of RNA polymerase II to a representative early promoter (thymidine kinase [TK]). Affinity purification experiments demonstrated that d3-10 ICP4 was not found in complexes with components of TFIID and mediator, suggesting that the defect in RNA polymerase II (Pol II) recruitment was the result of ablated interactions between d3-10 and TFIID and mediator. Complementation assays suggested that the N-terminal and C-terminal regions of ICP4 cooperate to mediate gene expression. The complementation was the result of the formation of more functional heterodimers, which restored the ability of the d3-10-containing molecules to interact with TFIID. Together, these studies suggest that the N terminus contains a true activation domain, mediating interactions with TFIID, mediator, and perhaps other transcription factors, and that the C terminus of the molecule contains activities that augment the functions of the activation domain.
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
authors.forename: Lauren M
authors.initials: LM
authors.lastname: Wagner
authors.forename: Avraham
authors.initials: A
authors.lastname: Bayer
authors.forename: Neal A
authors.initials: NA
authors.lastname: Deluca
mesh_terms: D000818:Animals; D002460:Cell Line; D004252:DNA Mutational Analysis; D017353:Gene Deletion; D015967:Gene Expression Regulation, Viral; D005816:Genetic Complementation Test; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D025941:Protein Interaction Mapping; D017384:Sequence Deletion; D018139:Simplexvirus / Q000235:genetics*; D035362:Transcription Factor TFIID / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D017874:Immediate-Early Proteins; D035362:Transcription Factor TFIID; C048925:herpes simplex virus, type 1 protein ICP4