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DNA methylation represses transcription in vivo

Nature Genetics volume 22, pages 203–206 (1999)Cite this article

Abstract

DNA in somatic tissue is characterized by a bimodal pattern of methylation, which is established in the animal through a series of developmental events1. In the mouse blastula, most DNA is unmethylated, but after implantation a wave of de novo methylation modifies most of the genome, excluding the majority of CpG islands, which are mainly associated with housekeeping genes. This genomic methylation pattern is broadly maintained during the life of the organism by maintenance methylation2, and generally correlates with gene expression. Experiments both in vitro3,4,5 and in vivo6,7,8,9 indicate that methylation inhibits transcription. It has not yet been possible, however, to determine the role of DNA methylation on specific sequences during normal development. Cis -acting regulatory elements and trans-acting factors appear to be involved in both stage- and tissue-specific demethylation processes10,11. Sp1-like elements have a key role in protecting the CpG island of Aprt (encoding adenine phosphoribosyltransferase) from de novo methylation, and when these elements are specifically mutated, the Aprt CpG island becomes methylated in transgenic mice12,13. We have now characterized an embryo-specific element from the CpG island sequence upstream of Aprt that can protect itself from de novo methylation in transgenic mice as well as reduce methylation of flanking sequences. We placed this element on a removable cassette adjacent to a human HBB (encoding β-globin) reporter and generated a transgene whose methylation pattern can be switched in vivo. Analysis of globin transcription in this system showed that methylation in cis inhibits gene expression in a variety of tissues, indicating that DNA modification may serve as a global genomic repressor.

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Figure 1: The IE prevents methylation.
Figure 2: IE removal switches methylation patterns in vivo.
Figure 3: DNA methylation represses background transcription.

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Acknowledgements

We thank E. Rand and T. Jakubowicz for help in preparing the manuscript and figures, and F.A. Asimakopoulos for help in the bisulfite analysis. This work was supported by grants from the NIH (H.C.), Council for Tobacco Research (H.C.), Israel Cancer Research fund (H.C.,Z.S.) and the Israel Ministry of Science (H.C.,Z.S.).

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Authors and Affiliations

  1. Department of Cellular Biochemistry, Hebrew University Medical School, Jerusalem, 91120, Israel

    Zahava Siegfried, Sharon Eden, Xu Feng, Ben-Zion Tsuberi & Howard Cedar

  2. Center for Neurobiology and Behavior, Columbia University Medical School, New York, 10032, USA, New York

    Monica Mendelsohn

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  1. Zahava Siegfried
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  2. Sharon Eden
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  3. Monica Mendelsohn
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  4. Xu Feng
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  6. Howard Cedar
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Correspondence to Howard Cedar.

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Siegfried, Z., Eden, S., Mendelsohn, M. et al. DNA methylation represses transcription in vivo. Nat Genet 22, 203–206 (1999). https://doi.org/10.1038/9727

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