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DNMT1 forms a complex with Rb, E2F1 and HDAC1 and represses transcription from E2F-responsive promoters

Nature Genetics volume 25, pages 338–342 (2000)Cite this article

Abstract

Methylation of CpG islands is associated with transcriptional silencing and the formation of nuclease-resistant chromatin structures enriched in hypoacetylated histones1,2,3. Methyl-CpG-binding proteins, such as MeCP2, provide a link between methylated DNA and hypoacetylated histones by recruiting histone deacetylase4,5, but the mechanisms establishing the methylation patterns themselves are unknown. Whether DNA methylation is always causal for the assembly of repressive chromatin or whether features of transcriptionally silent chromatin might target methyltransferase remains unresolved. Mammalian DNA methyltransferases show little sequence specificity in vitro, yet methylation can be targeted in vivo within chromosomes to repetitive elements6,7, centromeres8,9,10 and imprinted loci11. This targeting is frequently disrupted in tumour cells, resulting in the improper silencing of tumour-suppressor genes associated with CpG islands1,2. Here we show that the predominant mammalian DNA methyltransferase, DNMT1, co-purifies with the retinoblastoma (Rb) tumour suppressor gene product, E2F1, and HDAC1 and that DNMT1 cooperates with Rb to repress transcription from promoters containing E2F-binding sites. These results establish a link between DNA methylation, histone deacetylase and sequence-specific DNA binding activity, as well as a growth-regulatory pathway that is disrupted in nearly all cancer cells.

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Figure 1: Biochemical purification of a DNMT1 complex from HeLa cells.
Figure 2: DNMT1 and Rb interact directly both in vitro and in vivo.
Figure 3: DNMT1 and Rb repress transcription when artificially recruited to a promoter.
Figure 4: DNMT1 and Rb repress transcription from promoters containing E2F-binding sites.
Figure 5

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Acknowledgements

We thank A. Harel-Bellan for providing the E2F1 and Rb vectors, and J.-M. Zingg for advice with the DNMT trapping assay. S.A was supported by an EMBO fellowship.

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Author notes

  1. Keith D. Robertson

    Present address: Epigenetic Gene Regulation and Cancer/LRBGE, Bethesda, Maryland, USA

  2. Alan P. Wolffe

    Present address: Sangamo BioSciences Inc., Point Richmond Tech Center, Richmond, California, USA

  3. Keith D. Robertson and Slimane Ait-Si-Ali: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Molecular Embryology, NICHD, NIH, Bethesda, Maryland, USA

    Keith D. Robertson, Slimane Ait-Si-Ali, Tomoki Yokochi, Paul A. Wade, Peter L. Jones & Alan P. Wolffe

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Correspondence to Alan P. Wolffe.

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Robertson, K., Ait-Si-Ali, S., Yokochi, T. et al. DNMT1 forms a complex with Rb, E2F1 and HDAC1 and represses transcription from E2F-responsive promoters. Nat Genet 25, 338–342 (2000). https://doi.org/10.1038/77124

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