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PMID: 10673503 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Transient depletion of xDnmt1 leads to premature gene activation in Xenopus embryos.

Genes & development ·Vol. 14 ·No. 3 ·2000-02-01 ·Pages 313-27

Stancheva I, Meehan RR

Abstract

In Xenopus laevis zygotic transcription begins at the midblastula transition (MBT). Prior to this the genome is organized into chromatin that facilitates rapid cycles of DNA replication but not transcription. Here we demonstrate that DNA methylation contributes to the overall transcriptional silencing before MBT. Transient depletion of the maternal DNA methyltransferase (xDnmt1) by anti sense RNA during cleavage stages is associated with a decrease in the genomic 5-methyl-cytosine content and leads to the activation of zygotic transcription approximately two cell cycles earlier than normal. Hypomethylation allows the early expression of mesodermal marker genes such as Xbra, Cerberus, and Otx2, which are subsequently down-regulated during gastrulation of the xDnmt1-depleted embryos. The temporal switch in gene expression may account for the appearance of body plan defects that we observe. Loss of xDnmt1 can be rescued by the coinjection of mouse or human Dnmt1 protein. These results demonstrate that DNA methylation has a role in the regulation of immediately early genes in Xenopus at MBT.

MeSH Terms
5-Methylcytosine Animals Blotting, Northern Blotting, Western Cell Differentiation Cytosine/analogs & derivatives,chemistry DNA (Cytosine-5-)-Methyltransferase 1 DNA (Cytosine-5-)-Methyltransferases/chemistry,genetics,metabolism DNA Methylation Embryo, Nonmammalian Gastrula/metabolism Gene Expression Regulation, Developmental Gene Silencing Humans In Vitro Techniques Mice Microinjections RNA, Antisense/pharmacology Reverse Transcriptase Polymerase Chain Reaction T-Box Domain Proteins/metabolism Transcriptional Activation Xenopus Proteins Xenopus laevis
Chemicals
RNA, Antisense T-Box Domain Proteins TBXT protein, Xenopus Xenopus Proteins 5-Methylcytosine Cytosine DNA (Cytosine-5-)-Methyltransferase 1 DNA (Cytosine-5-)-Methyltransferases DNMT1 protein, human Dnmt1 protein, mouse
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Stancheva I
Genes and Development Group, Department of Biomedical Sciences, University of Edinburgh, Edinburgh EH8 9XD Scotland, UK.
Meehan R R
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
Published
2000-02-01
Pages
313-27
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC316362
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source PMC Full Text View journal details
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