Chemical synthesis of 5-azacytidine nucleotides and preparation of tRNAs containing 5-azacytidine in its 3'-terminus.
5-azacytidine-5'-triphosphate prepared from 5-azacytidine by chemical phosphorylation is a substrate for AMP (CMP) tRNA nucleotidyl transferase from yeast. tRNAsPhe from yeast containing 5-azacytidine in their 3'-termini were prepared enzymatically. tRNAPhe-Cpn5CpA and tRNAPhe-n5Cpn5CpA can be aminoacylated by phenylalanyl-tRNA synthetase from yeast and they are active in the poly(U)-dependent synthesis of poly(Phe) on E. coli ribosomes. The decomposition of 5-azacytidine via hydrolysis of the triazine ring is significantly accelerated by a phosphate group on the 5'-position of the nucleotide. After the incorporation of 5-azacytidine-5'-phosphate into a polynucleotide chain the rate of hydrolysis of the triazine ring decreases considerably.
D001374:Azacitidine / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D003568:Cytidine Monophosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D003570:Cytidine Triphosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D003597:Cytosine Nucleotides; D004926:Escherichia coli / Q000378:metabolism; D007202:Indicators and Reagents; D007700:Kinetics; D010652:Phenylalanine-tRNA Ligase / Q000378:metabolism; D010766:Phosphorylation; D014176:Protein Biosynthesis; D012316:RNA Nucleotidyltransferases / Q000378:metabolism; D012343:RNA, Transfer / Q000138:chemical synthesis* / Q000235:genetics; D012270:Ribosomes / Q000378:metabolism
D003597:Cytosine Nucleotides; D007202:Indicators and Reagents; C012201:5'-azacytidine 5'-monophosphate; C012202:5'-azacytidine 5'-triphosphate; D003570:Cytidine Triphosphate; D012343:RNA, Transfer; D012316:RNA Nucleotidyltransferases; C023198:tRNA nucleotidyltransferase; D010652:Phenylalanine-tRNA Ligase; D003568:Cytidine Monophosphate; D001374:Azacitidine