Deoxyuridine triphosphate

Fig. lA. Anabolic and catabolic pathways of 5-FU. DPD dihydropyrimidine dehydrogenase, DP di-hydropyrimidinase, pUP beta-ureidopropionase, UP uridine phosphorylase, OPRT orotate phospho-ribosyl transferase, UK uridine kinase, TP thymidine phosphorylase, TK thymidine kinase, RNR ribonucleotide reductase. The three active metabolites (shown in rectangles) are FdUMP (5-fluoro-2 -deoxyuridine 5 -monophosphate) inhibiting TS (thymidylate synthase), and FUTP (5-fluorouridine 5 -triphosphate) and FdUTP (5-fluoro 2 -deoxyuridine 5 -triphosphate) interfering with RNA and DNA, respectively. 

Fig. 31. Connection of 5-(3"-aminopropynyl)-2 -deoxyuridine-5 -triphosphate to the succinimide active ester arm of a ruthenium polypyridyl complex.

The full synthesis of two Fc-labeled bases 5-(3-ferrocenecarboxamidopropenyl-l) 2 -deoxyuridine 5 -triphosphate (Fcl-dUTP) and 5-(3-ferroceneacetamidopropenyl-... 

Biotin-1 l-deoxyuridine-5 -triphosphate (BiodUTP) and reagents for its incorporation into DNA by nick translation are obtained commercially. The products from Bethesda Research Laboratories (BRL, Gaithersbui, MD) are acceptable. BRL now provides a prepackaged kit (BioNick) containing necessary supplies and employing biotin-14-dATP as the source of biotin. The concentration of the resulting biotinylated DNA is determined by the histochemical method for biotin (below). Adequate instructions are provided with these kits. 

The triphosphate of 5-formyldUTP (111) has been prepared and its incorporation by several DNA polymerases has been studied. It was found to be incorporated well in place of TTP and could also substitute dCTP to some extent. The preparation of 5-azidoUTP (112) from UTP has been described. The analogue was found to be an inhibitor of CPasell. 2 -Deoxyuridine-5 -triphosphate labelled at the C5 position with the dye methylene blue has been prepared and incorporated into oligonucleotides or DNA fragments by tailing with terminal transferase. " ... 

Azido-2 -deoxyuridine 5 -triphosphate (5-N3-dUTP) has been enzymically converted by E. coli DNA polymerase (in the dark) to a photoreactive DNA which was then used to photolabel the lac repressor (230). Alternatively, when 5-N3-dUTP was irradiated in the presence of DNA polymerase, it functioned as a photoaffinity label of the enzyme (231). The syntheses of several photoreactive 3 -arylazido derivatives of ATP (232) as well as of 9-(3 -azido-3 -deoxy-/3-D-xylofuranosyl)adenosine and -guanosine 5 -triphosphates (233) have been described. 

Mertes investigated the coupling of the 5-mercuriuridines with styrenes in aqueous media resulting in alkylation of the uracil nucleotides. Carbon alkylation of C-5 on the uracil ring in the ribo- and deoxyri-bonucleosides and nucleotides was obtained in high yields by this method. A similar reaction was used by Langer et al. in the synthesis of 5-(3-amino)allyluridine and deoxyuridine-5 -triphosphates (AA-UTP and AA-dUTP) (6.12) ... 

Many cells can synthesize deoxyuridine 5 -triphosphate (dUTP). Can dUTP be used as a substrate for DNA polymerase If so, with which base will uracil pair in newly replicated DNA ... 

The de novo pathway to 2 -deoxythymidine monophosphate (dTMP) synthesis first requires the use of dUMP (2 -deoxyuridine-5 -monophosphate) from the metabolism of either UDP or CDP (cytidine diphosphate). The hydrolysis of dUTP (2 -deoxyuridine-5 -triphosphate) to dUMP and subsequent methylation at C-5 by the action of thymidylate synthase, using A, A i°-methylenetetrahydrofolate (THF) as the methyl donor, generate dTMP (Figure 6.54). The latter is subsequently phosphorylated to deoxy-thymidine triphosphate (dTTP) used in DNA synthesis and repair. 

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