Inhibition of herpes simplex virus 1 gene expression by designer zinc-finger transcription factors.

TitleInhibition of herpes simplex virus 1 gene expression by designer zinc-finger transcription factors.
Publication TypeJournal Article
Year of Publication2003
AuthorsPapworth, M, Moore, M, Isalan, M, Minczuk, M, Choo, Y, Klug, A
JournalProc Natl Acad Sci U S A
Volume100
Issue4
Pagination1621-6
Date Published2003 Feb 18
ISSN0027-8424
KeywordsAnimals, Base Sequence, Chloramphenicol O-Acetyltransferase, COS Cells, DNA Primers, Fluorescent Antibody Technique, Gene Expression Regulation, Viral, Genes, Viral, HeLa Cells, Humans, Simplexvirus, Transcription Factors, Zinc Fingers
Abstract

The herpes simplex virus 1 (HSV-1) replicative cycle begins by binding of the viral activator, VP16, to a set of sequences in the immediate-early (IE) gene promoters. With the aim of inhibiting this cycle, we have constructed a number of synthetic zinc-finger DNA-binding peptides by using recently reported methods. Peptides containing either three or six fingers, targeted to a viral promoter, were engineered as fusions with a KOX-1 transcription repression domain. These proteins bound to the HSV-1 IE175k (ICP4) promoter, in vitro, with nanomolar or subnanomolar binding affinity. However, in a chloramphenicol acetyltransferase reporter system, only the six-finger protein was found to repress VP16-activated transcription significantly. Thus the longer array of zinc fingers is required to compete successfully against VP16, one of the most powerful natural activators known. We found that the HSV-1 replication cycle can be partially repressed by the six-finger peptide with the viral titer reduced by 90%.

DOI10.1073/pnas.252773399
Alternate JournalProc. Natl. Acad. Sci. U.S.A.
Citation Key10.1073/pnas.252773399
PubMed ID12574501
PubMed Central IDPMC149882