Noyon Caroline, Roumeguère Thierry, Delporte Cédric, Dufour Damien, Cortese Melissa, Desmet Jean-Marc, Lelubre Christophe, Rousseau Alexandre, Poelvoorde Philippe, Nève Jean, Vanhamme Luc, Boudjeltia Karim Zouaoui, Van Antwerpen Pierre
Laboratory of Organic Pharmaceutical Chemistry, Faculty of Pharmacy, Université Libre de Bruxelles, 1050, Brussels, Belgium.
Department of Urology, Erasme Hospital, Université Libre de Bruxelles, 1070, Brussels, Belgium.
Mol Cell Biochem. 2017 May;429(1-2):59-71. doi: 10.1007/s11010-016-2936-2. Epub 2017 Jan 10.
Myeloperoxidase (MPO) is able to promote several kinds of damage and is involved in mechanisms leading to various diseases such as atherosclerosis or cancers. An example of these damages is the chlorination of nucleic acids, which is considered as a specific marker of the MPO activity. Since 5-chlorocytidine has been recently shown in healthy donor plasmas, this study aimed at discovering if these circulating modified nucleosides could be incorporated into RNA and DNA and if their presence impacts the ability of enzymes involved in the incorporation, transcription, and translation processes. Experimentations, which were carried out in vitro with endothelial and prostatic cells, showed a large penetration of all chloronucleosides but an exclusive incorporation of 5-chlorocytidine into RNA. However, no incorporation into DNA was observed. This specific incorporation is accompanied by an important reduction of translation yield. Although, in vitro, DNA polymerase processed in the presence of chloronucleosides but more slowly than in control conditions, ribonucleotide reductase could not reduce chloronucleotides prior to the replication. This reduction seems to be a limiting step, protecting DNA from chloronucleoside incorporation. This study shows the capacity of transcription enzyme to specifically incorporate 5-chlorocytidine into RNA and the loss of capacity-complete or partial-of different enzymes, involved in replication, transcription or translation, in the presence of chloronucleosides. Questions remain about the long-term impact of such specific incorporation in the RNA and such decrease of protein production on the cell viability and function.
髓过氧化物酶(MPO)能够引发多种损伤,并参与导致动脉粥样硬化或癌症等各种疾病的机制。这些损伤的一个例子是核酸的氯化作用,这被认为是MPO活性的一种特异性标志物。由于最近在健康供体血浆中发现了5-氯胞苷,本研究旨在探究这些循环中的修饰核苷是否能够掺入RNA和DNA,以及它们的存在是否会影响参与掺入、转录和翻译过程的酶的活性。在内皮细胞和前列腺细胞中进行的体外实验表明,所有氯核苷都有大量渗透,但只有5-氯胞苷特异性地掺入RNA。然而,未观察到其掺入DNA。这种特异性掺入伴随着翻译产量的显著降低。虽然在体外,DNA聚合酶在氯核苷存在的情况下仍能发挥作用,但比对照条件下要慢,而核糖核苷酸还原酶在复制前无法还原氯核苷酸。这种还原似乎是一个限制步骤,可保护DNA不被氯核苷掺入。本研究表明转录酶能够将5-氯胞苷特异性地掺入RNA,并且在氯核苷存在的情况下,参与复制、转录或翻译的不同酶的活性会全部或部分丧失。关于这种特异性掺入RNA以及蛋白质产量下降对细胞活力和功能的长期影响,仍存在疑问。