DOI: 10.1083/jcb.200610062. DNMT1 but not its interaction with the replication machinery is required for maintenance of DNA methylation in human cells

DNMT1 but not its interaction with the replication machinery is required for maintenance of DNA methylation in human cells

10.1083/jcb.200610062

Crossref journal-article

Rockefeller University Press

The Journal of Cell Biology (291)

Abstract

DNA methylation plays a central role in the epigenetic regulation of gene expression in vertebrates. Genetic and biochemical data indicated that DNA methyltransferase 1 (Dnmt1) is indispensable for the maintenance of DNA methylation patterns in mice, but targeting of the DNMT1 locus in human HCT116 tumor cells had only minor effects on genomic methylation and cell viability. In this study, we identified an alternative splicing in these cells that bypasses the disrupting selective marker and results in a catalytically active DNMT1 protein lacking the proliferating cell nuclear antigen–binding domain required for association with the replication machinery. Using a mechanism-based trapping assay, we show that this truncated DNMT1 protein displays only twofold reduced postreplicative DNA methylation maintenance activity in vivo. RNA interference–mediated knockdown of this truncated DNMT1 results in global genomic hypomethylation and cell death. These results indicate that DNMT1 is essential in mouse and human cells, but direct coupling of the replication of genetic and epigenetic information is not strictly required.

Bibliography

Spada, F., Haemmer, A., Kuch, D., Rothbauer, U., Schermelleh, L., Kremmer, E., Carell, T., LaÌngst, G., & Leonhardt, H. (2007). DNMT1 but not its interaction with the replication machinery is required for maintenance of DNA methylation in human cells. The Journal of Cell Biology, 176(5), 565â571.

Authors 9

Fabio Spada (first)
Andrea Haemmer (additional)
David Kuch (additional)
Ulrich Rothbauer (additional)
Lothar Schermelleh (additional)
Elisabeth Kremmer (additional)
Thomas Carell (additional)
Gernot Längst (additional)
Heinrich Leonhardt (additional)

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Dates

Type	When
Created	18 years, 6 months ago (Feb. 20, 2007, 11:24 p.m.)
Deposited	2 years, 1 month ago (July 22, 2023, 4:19 p.m.)
Indexed	1 month ago (Aug. 2, 2025, 12:27 a.m.)
Issued	18 years, 6 months ago (Feb. 20, 2007)
Published	18 years, 6 months ago (Feb. 20, 2007)
Published Online	18 years, 6 months ago (Feb. 20, 2007)
Published Print	18 years, 6 months ago (Feb. 26, 2007)

Funders 0

None

BibTeX

@article{Spada_2007, title={DNMT1 but not its interaction with the replication machinery is required for maintenance of DNA methylation in human cells}, volume={176}, ISSN={0021-9525}, url={http://dx.doi.org/10.1083/jcb.200610062}, DOI={10.1083/jcb.200610062}, number={5}, journal={The Journal of Cell Biology}, publisher={Rockefeller University Press}, author={Spada, Fabio and Haemmer, Andrea and Kuch, David and Rothbauer, Ulrich and Schermelleh, Lothar and Kremmer, Elisabeth and Carell, Thomas and Längst, Gernot and Leonhardt, Heinrich}, year={2007}, month=feb, pages={565–571} }

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