Light-Activatable TET-Dioxygenases Reveal Dynamics of 5-Methylcytosine Oxidation and Transcriptome Reorganization
| Autor: | Paul Czodrowski, Benjamin Buchmuller, Sascha Jung, Jan Wolffgramm, Álvaro Muñoz-López, Daniel Summerer, Shubhendu Palei |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
010402 general chemistry
01 natural sciences Biochemistry Catalysis Dioxygenases Transcriptome chemistry.chemical_compound Colloid and Surface Chemistry Transcriptional regulation Humans Epigenetics General Chemistry Epigenome 0104 chemical sciences Chromatin Cell biology 5-Methylcytosine DNA demethylation chemistry Oxidation-Reduction DNA |
| Zdroj: | Journal of the American Chemical Society |
| DOI: | 10.17877/de290r-21201 |
| Popis: | Ten-eleven-translocation (TET) dioxygenases catalyze the oxidation of 5-methylcytosine (5mC), the central epigenetic regulator of mammalian DNA. This activity dy- namically reshapes epigenome and transcriptome by deposit- ing oxidized 5mC derivatives, and initiating active DNA de- methylation. However, studying this dynamic is hampered by the inability to selectively activate individual TETs with tem- poral control in cells. We report activation of TETs in mam- malian cells by incorporation of genetically encoded 4,5- dimethoxy-2-nitrobenzyl-L-serine as transient active site block, and its subsequent deprotection with light. Our ap- proach enables precise insights into the impact of cancer- associated TET2 mutations on the kinetics of TET2 catalysis in vivo, and allows time-resolved monitoring of target gene activation and transcriptome reorganization. This sets a basis for dissecting the order and kinetics of chromatin-associated events triggered by TET catalysis, ranging from DNA de- methylation to chromatin and transcription regulation. |
| Databáze: | OpenAIRE |
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