Spada, Fabio and Haemmer, Andrea and Kuch, David and Rothbauer, Ulrich and Schermelleh, Lothar and Kremmer, Elisabeth and Carell, Thomas and Laengst, Gernot and Leonhardt, Heinrich (2007) DNMT1 but not its interaction with the replication machinery is required for maintenance of DNA methylation in human cells. JOURNAL OF CELL BIOLOGY, 176 (5). pp. 565-571. ISSN 0021-9525, 1540-8140
Full text not available from this repository. (Request a copy)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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | HUMAN CANCER-CELLS; EMBRYONIC STEM-CELLS; CYTOSINE-5 METHYLTRANSFERASE; CPG METHYLATION; DE-NOVO; HYPOMETHYLATION; PURIFICATION; COMPLEX; SITES; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III > Prof. Dr. Gernot Längst |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 22 Dec 2020 11:56 |
| Last Modified: | 22 Dec 2020 11:56 |
| URI: | https://pred.uni-regensburg.de/id/eprint/33243 |
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