Investigating and controlling mRNA via chemo-enzymatic modifications

Methylation of DNA, RNA and proteins constitutes a major regulatory mechanism of biological processes. In nature, S‑adenosyl-l-methionine (AdoMet) is the cosubstrate of most methyltransferases (MTases) and the main methyl source. Synthetic AdoMet analogues are also accepted by a number of promiscuous MTases or engineered variants.
In this talk, I will show how AdoMet analogs in combination with suitable MTases can be used to transfer clickable or photo-cleavable (PC) groups to DNA, RNA and the mRNA 5' cap.^[1] Post-synthetic bioconjugation makes MTase target sites in mRNA detectable via next generation sequencing.^[2] Light-induced removal provides methodology for site-specific “writing” and “erasing” marks on long DNA or RNA at natural MTase sites.
As a result of protein engineering of methionine adenosyltransferase (MAT), enzymatic in situ generation of AdoMet analogs with PC-groups from their amino acid precursors is now possible.^[3] The enzymatic cascade accepting photocaging groups bears potential for future cellular applications.

Meeting ID: 683 0972 8656
Passcode: 974255