Synthesis of Ribonucleoside 5'-Triphosphates Derived from 2-Thiocytidine, 4-Thiouridine, and 5-Bromocytidine
نویسنده
چکیده
The synthesis of 5'-triphosphates of 4-thiouridine, 2-thiocytidine, 5-bromocytidine, and [6N]adenosine is described. All compounds are characterized by chromatographic and spectral data. Phosphorylation of corresponding nucleotides was performed using phosphoryl chloride and con densation with pyrophosphate. Intermediates were not isolated. The final products were purified by chromatography and characterized by 3 1 P NMR spectroscopy. The nucleoside triphosphates are substrates for RNA polymerase from bacteriophage T7 and were used for enzymatic synthesis of modified and isotopically labelled RNA.
منابع مشابه
Phosphorylation of uridine and cytidine nucleoside analogs by two human uridine-cytidine kinases.
Uridine-cytidine kinases (UCK) have important roles for the phosphorylation of nucleoside analogs that are being investigated for possible use in chemotherapy of cancer. We have cloned the cDNA of two human UCKs. The approximately 30-kDa proteins, named UCK1 and UCK2, were expressed in Escherichia coli and shown to catalyze the phosphorylation of Urd and Cyd. The enzymes did not phosphorylate d...
متن کاملFormation of thiolated nucleosides present in tRNA from Salmonella enterica serovar Typhimurium occurs in two principally distinct pathways.
tRNA from Salmonella enterica serovar Typhimurium contains five thiolated nucleosides, 2-thiocytidine (s(2)C), 4-thiouridine (s(4)U), 5-methylaminomethyl-2-thiouridine (mnm(5)s(2)U), 5-carboxymethylaminomethyl-2-thiouridine (cmnm(5)s(2)U), and N-6-(4-hydroxyisopentenyl)-2-methylthioadenosine (ms(2)io(6)A). The levels of all of them are significantly reduced in cells with a mutated iscS gene, wh...
متن کاملThe cysteine desulfurase IscS is required for synthesis of all five thiolated nucleosides present in tRNA from Salmonella enterica serovar typhimurium.
Deficiency of a modified nucleoside in tRNA often mediates suppression of +1 frameshift mutations. In Salmonella enterica serovar Typhimurium strain TR970 (hisC3737), which requires histidine for growth, a potential +1 frameshifting site, CCC-CAA-UAA, exists within the frameshifting window created by insertion of a C in the hisC gene. This site may be suppressed by peptidyl-tRNAProcmo5UGG (cmo(...
متن کاملIn vitro transcription of labeled RNA: synthesis, capping, and substitution.
All in vitro RNA transcription reactions involve the use of common components: a template, phage RNA polymerase, and ribonucleoside triphosphates (rNTPs). The concentration of rNTPs is a crucial variable that is manipulated to determine the specific activities of labeled RNAs. This protocol describes methods used to synthesize RNAs of low specific activity (trace labeled), medium specific activ...
متن کاملNew NTP analogs: the synthesis of 4′-thioUTP and 4′-thioCTP and their utility for SELEX
The synthesis of the triphosphates of 4'-thiouridine and 4'-thiocytidine, 4'-thioUTP (7; thioUTP) and 4'-thioCTP (8; thioCTP), and their utility for SELEX (systematic evolution of ligands by exponential enrichment) is described. The new nucleoside triphosphate (NTP) analogs 7 and 8 were prepared from appropriately protected 4'-thiouridine and -cytidine derivatives using the one-pot method repor...
متن کامل