title: Mutational analysis of a transcriptional activation region of the VP16 protein of herpes simplex virus.
abstract: The VP16 protein of herpes simplex virus is a potent transcriptional activator of the viral immediate early genes. The transcriptional activation region of VP16 can be divided into two functional subregions, here designated VP16N (comprising amino acids 413-456) and VP16C (amino acids 450-490). Assays of VP16C mutants resulting from both random and alanine-scanning mutagenesis indicated that the sidechains of three phenylalanines (at positions 473, 475 and 479) and one acidic residue (glutamate 476) are important for transcriptional activation. Aromatic and bulky hydrophobic amino acids were effective substitutes for each of the three Phe residues, whereas replacement with smaller or polar amino acids resulted in loss of transcriptional function. In contrast, many changes were tolerated for Glu476, including bulky hydrophobic and basic amino acids, indicating that the negative charge at this position contributes little to the function of this subregion. Similar relative activities for most of the mutants were observed in yeast and in mammalian cells, indicating that the structural requirements for this activation region are comparable in these two species. These results reinforce the hypothesis that bulky hydrophobic residues, not acidic residues, are most critical for the activity of this 'acidic' transcriptional activation region.
authors.affiliation: Department of Biochemistry, Michigan State University, East Lansing, MI 48824-1319, USA.
authors.forename: S M
authors.initials: SM
authors.lastname: Sullivan
authors.forename: P J
authors.initials: PJ
authors.lastname: Horn
authors.forename: V A
authors.initials: VA
authors.lastname: Olson
authors.forename: A H
authors.initials: AH
authors.lastname: Koop
authors.forename: W
authors.initials: W
authors.lastname: Niu
authors.forename: R H
authors.initials: RH
authors.lastname: Ebright
authors.forename: S J
authors.initials: SJ
authors.lastname: Triezenberg
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics*; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D018139:Simplexvirus / Q000235:genetics*; D015533:Transcriptional Activation / Q000235:genetics*
chemical_list: D004279:DNA, Viral; D017405:Herpes Simplex Virus Protein Vmw65