Chimeric DNA methyltransferases target DNA methylation to specific DNA sequences and repress expression of target genes.

TitleChimeric DNA methyltransferases target DNA methylation to specific DNA sequences and repress expression of target genes.
Publication TypeJournal Article
Year of Publication2007
AuthorsLi, F, Papworth, M, Minczuk, M, Rohde, C, Zhang, Y, Ragozin, S, Jeltsch, A
JournalNucleic Acids Res
Volume35
Issue1
Pagination100-12
Date Published2007
ISSN1362-4962
KeywordsAnimals, Base Sequence, Binding Sites, Cell Line, DNA, DNA (Cytosine-5-)-Methyltransferases, DNA Methylation, Gene Silencing, Genes, ras, Herpesvirus 1, Human, Humans, Immediate-Early Proteins, Mice, Promoter Regions, Genetic, Protein Structure, Tertiary, Recombinant Fusion Proteins, Transcription Factors, Viral Proteins, Zinc Fingers
Abstract

Gene silencing by targeted DNA methylation has potential applications in basic research and therapy. To establish targeted methylation in human cell lines, the catalytic domains (CDs) of mouse Dnmt3a and Dnmt3b DNA methyltransferases (MTases) were fused to different DNA binding domains (DBD) of GAL4 and an engineered Cys2His2 zinc finger domain. We demonstrated that (i) Dense DNA methylation can be targeted to specific regions in gene promoters using chimeric DNA MTases. (ii) Site-specific methylation leads to repression of genes controlled by various cellular or viral promoters. (iii) Mutations affecting any of the DBD, MTase or target DNA sequences reduce targeted methylation and gene silencing. (iv) Targeted DNA methylation is effective in repressing Herpes Simplex Virus type 1 (HSV-1) infection in cell culture with the viral titer reduced by at least 18-fold in the presence of an MTase fused to an engineered zinc finger DBD, which binds a single site in the promoter of HSV-1 gene IE175k. In short, we show here that it is possible to direct DNA MTase activity to predetermined sites in DNA, achieve targeted gene silencing in mammalian cell lines and interfere with HSV-1 propagation.

DOI10.1093/nar/gkl1035
Alternate JournalNucleic Acids Res.
Citation Key10.1093/nar/gkl1035
PubMed ID17151075
PubMed Central IDPMC1761428