Dr Jekyll and Mr Hyde: a strange case of 5-ethynyl-2 '-deoxyuridine and 5-ethynyl-2 '- deoxycytidine

0458684 - ÚOCHB 2017 RIV GB eng J - Journal Article
Ligasová, A. - Liboska, Radek - Friedecký, D. - Mičová, K. - Adam, T. - Oždian, T. - Rosenberg, Ivan - Koberna, K.
Dr Jekyll and Mr Hyde: a strange case of 5-ethynyl-2 '-deoxyuridine and 5-ethynyl-2 '- deoxycytidine.
Open Biology. Roč. 6, č. 1 (2016), č. článku 150172. E-ISSN 2046-2441
R&D Projects: GA MZd NV15-31604A
Institutional support: RVO:61388963
Keywords : cytidine deaminase * dCMP deaminase * 5-ethynyl-2 '-deoxyuridine * 5-ethynyl-2 '-deoxycytidine * DNA replication
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 3.481, year: 2016
http://rsob.royalsocietypublishing.org/content/6/1/150172

5-Ethynyl-2 '-deoxyuridine (EdU) and 5-ethynyl-2 '-deoxycytidine (EdC) are mainly used as markers of cellular replicational activity. Although EdU is employed as a replicational marker more frequently than EdC, its cytotoxicity is commonly much higher than the toxicity of EdC. To reveal the reason of the lower cytotoxicity of EdC, we performed a DNA analysis of five EdC-treated human cell lines. Surprisingly, not a single one of the tested cell lines contained a detectable amount of EdC in their DNA. Instead, the DNA of all the cell lines contained EdU. The content of incorporated EdU differed in particular cells and EdC-related cytotoxicity was directly proportional to the content of EdU. The results of experiments with the targeted inhibition of the cytidine deaminase (CDD) and dCMP deaminase activities indicated that the dominant role in the conversion pathway of EdC to EdUTP is played by CDD in HeLa cells. Our results also showed that the deamination itself was not able to effectively prevent the conversion of EdC to EdCTP, the conversion of EdC to EdCTP occurs with much lesser effectivity than the conversion of EdU to EdUTP and the EdCTP is not effectively recognized by the replication complex as a substrate for the synthesis of nuclear DNA.
Permanent Link: http://hdl.handle.net/11104/0258938