Mutational analysis of a transcriptional activation region of the VP16 protein of herpes simplex virus

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). Assa...

Full description

Saved in:
Bibliographic Details
Published in:Nucleic acids research 1998-10, Vol.26 (19), p.4487-4496
Main Authors: Sullivan, Susan M., Horn, Peter J., Olson, Victoria A., Triezenberg, Steven J., Koop, Allen H., Niu, Wei, Ebright, Richard H.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
DNA Mutational Analysis
DNA, Viral - analysis
DNA, Viral - genetics
Herpes simplex virus
Herpes Simplex Virus Protein Vmw65 - genetics
Molecular Sequence Data
Mutation
Simplexvirus - genetics
Transcriptional Activation - genetics
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0305-1048
1362-4962
1362-4962
DOI:10.1093/nar/26.19.4487