Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Flemingovo nam. 2, 16610, Prague 6, Czech Republic.
Department of Organic Chemistry, Faculty of Science, Charles University, Hlavova 8, CZ-12843, Prague 2, Czech Republic.
Chemistry. 2022 Jun 1;28(31):e202200911. doi: 10.1002/chem.202200911. Epub 2022 Apr 13.
5-(β-d-Glucopyranosyloxymethyl)-2'-deoxyuridine and -cytidine 5'-O-triphosphates were prepared and used for polymerase-mediated (primer extension or PCR) synthesis of DNA containing glucosylated 5-hydroxymethyluracil (5hmU) or 5-hydroxymethyluracil (5hmC). The presence of any glucosylated pyrimidines fully protected DNA from cleavage by type II restriction endonucleases. On the other hand, while the presence of glucosylated 5hmU completely inhibited transcription by bacterial (Escherichia coli) RNA polymerase, the DNA containing the corresponding glucosylated 5hmC allowed a similar level of transcription as natural DNA. This suggests different roles of these hypermodified bases in the epigenetic regulation of transcription in bacteriophages or kinetoplastid parasites. Consequently, enzymatic glucosylation of 5hmC-containing DNA can be used for tuning of transcription activity.
5-(β-d-葡萄糖基氧甲基)-2'-脱氧尿苷和-胞苷 5'-O-三磷酸酯被制备并用于聚合酶介导的（引物延伸或 PCR）含有葡萄糖基化 5-羟甲基尿嘧啶（5hmU）或 5-羟甲基尿嘧啶（5hmC）的 DNA 的合成。任何葡萄糖基化嘧啶的存在都能完全保护 DNA 免受 II 型限制内切酶的切割。另一方面，虽然含有葡萄糖基化 5hmU 的 DNA 完全抑制了细菌（大肠杆菌）RNA 聚合酶的转录，但含有相应葡萄糖基化 5hmC 的 DNA 允许与天然 DNA 相似水平的转录。这表明这些超修饰碱基在噬菌体或动基体寄生虫中转录的表观遗传调控中发挥不同的作用。因此，含 5hmC 的 DNA 的酶促葡萄糖基化可用于调节转录活性。
