Abstract
Regulation of cell fate requires the establishment and erasure of 5-methylcytosine(5mC)in genomic DNA.The formation of 5mC is achieved by DNA cytosine methyltransferases(DNMTs),whereas the removal of 5mC can be accomplished by various pathways.Aside from ten-eleven translocation(TET)-mediated ox-idation of 5mC followed by thymine DNA glycosylase(TDG)-initiated base excision repair(BER),the di-rect deformylation of 5-formylcytosine(5fC)and decarboxylation of 5-carboxylcytosine(5caC)have also been discovered as the novel DNA demethylation pathways.Although these novel demethylation path-ways have been identified in stem cells and somatic cells,their precise roles in regulating cell fate re-main unclear.Here,we differentiate mouse embryonic stem cells(mESCs)into mouse embryoid bodies(mEBs),followed by further differentiation into mouse neural stem cells(mNSCs)and finally into mouse neurons(mNeurons).During this sequential differentiation process,we employ probe molecules,namely 2'-fluorinated 5-formylcytidine(F-5fC)and 2'-fluorinated 5-carboxyldeoxycytidine(F-5caC),for metabolic labeling.The results of mass spectrometry(MS)analysis demonstrate the deformylation and decarboxy-lation activities are progressively decreased and increased respectively during differentiation process,and this opposite demethylation tendency is not associated with DNMTs and TETs.
维普
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