Inhibition of herpes simplex virus 1 gene expression by designer zinc-finger transcription factors.
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%.
Papworth
Monika
M
Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom. map@mrc-lmb.cam.ac.uk
Moore
Michael
M
Isalan
Mark
M
Minczuk
Michal
M
Choo
Yen
Y
Klug
Aaron
A
D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D017931:DNA Primers; D005455:Fluorescent Antibody Technique; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D005814:Genes, Viral; D006367:HeLa Cells; D006801:Humans; D018139:Simplexvirus / Q000235:genetics*; D014157:Transcription Factors / Q000737:chemistry / Q000502:physiology*; D016335:Zinc Fingers
D017931:DNA Primers; D014157:Transcription Factors; D015500:Chloramphenicol O-Acetyltransferase