Authors: | Palei, Shubhendu Buchmuller, Benjamin Wolffgramm, Jan Muñoz-Lopez, Álvaro Jung, Sascha Czodrowski, Paul Summerer, Daniel |
Title: | Light-activatable TET-dioxygenases reveal dynamics of 5-Methylcytosine oxidation and transcriptome reorganization |
Language (ISO): | en |
Abstract: | 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. |
URI: | http://hdl.handle.net/2003/40993 http://dx.doi.org/10.17877/DE290R-22842 |
Issue Date: | 2020-04-14 |
Appears in Collections: | Chemische Biologie |
Files in This Item:
File | Description | Size | Format | |
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200319_ShP_Caged TET_nonmarked.pdf | 8.44 MB | Adobe PDF | View/Open |
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