An embryonic demethylation mechanism involving binding of transcription factors to replicating DNA

Koichi Matsuo, John Silke, Oleg Georgiev, Philippe Marti, Natalia Giovannini, Duri Rungger

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

In vertebrates, transcriptionally active promoters are undermethylated. Since the transcription factor Sp1, and more recently NF-κB, have been implicated in the demethylation process, we examined the effect of transcription factors on demethylation by injecting in vitro methylated plasmid DNA into Xenopus fertilized eggs. We found that various transactivation domains, including a strong acidic activation domain from the viral protein VP16, can enhance demethylation of a promoter region when fused to a DNA binding domain which recognizes the promoter. Furthermore, demethylation occurs only after the midblastula transition, when the general transcription machinery of the host embryo becomes available. Nevertheless, transcription factor binding need not be followed by actual transcription, since demethylation is not blocked by α-amanitin treatment. Finally, replication of the target DNA is a prerequisite for efficient demethylation since only plasmids that carry the bovine papilloma virus sequences which support plasmid replication after the midblastula transition are demethylated. No demethylation is detectable in the oocyte system where DNA is not replicated. These results suggest that, in the Xenopus embryo, promoters for which transcription factors are available are demethylated by a replication-dependent, possibly passive mechanism.

Original languageEnglish
Pages (from-to)1446-1453
Number of pages8
JournalEMBO Journal
Volume17
Issue number5
DOIs
Publication statusPublished - 1998 Mar 2
Externally publishedYes

Fingerprint

Plasmids
Transcription Factors
Xenopus
DNA
Transcription
Embryonic Structures
Amanitins
Sp1 Transcription Factor
Zygote
Papilloma
Viral Proteins
DNA Replication
Genetic Promoter Regions
Transcriptional Activation
Oocytes
Vertebrates
Viruses
Machinery
Chemical activation
In Vitro Techniques

Keywords

  • Bovine papilloma virus (PBV)
  • Gal4 fusion proteins
  • Sp1
  • Xenopus embryos
  • Xenopus oocytes

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

An embryonic demethylation mechanism involving binding of transcription factors to replicating DNA. / Matsuo, Koichi; Silke, John; Georgiev, Oleg; Marti, Philippe; Giovannini, Natalia; Rungger, Duri.

In: EMBO Journal, Vol. 17, No. 5, 02.03.1998, p. 1446-1453.

Research output: Contribution to journalArticle

Matsuo, Koichi ; Silke, John ; Georgiev, Oleg ; Marti, Philippe ; Giovannini, Natalia ; Rungger, Duri. / An embryonic demethylation mechanism involving binding of transcription factors to replicating DNA. In: EMBO Journal. 1998 ; Vol. 17, No. 5. pp. 1446-1453.
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