Preparation of Deoxy-5-hydroxymethylcytidine Triphosphate and its Monoglucosyl derivative

Abstract

The deoxyribonucleic acids of the Escherichia coli bacteriophage T2, T4, and T6 contain the pyrimidine nucleotide deoxy- 5-hydroxymethylcytidylic acid and derivatives with one or two glucosyl groups substituted on the 5-hydroxymethyl group (1-3). In order to study the possible role of the triphosphates of these nucleotides in the biosynthesis of the phage deoxyribonucleic acid, we have prepared the triphosphates of the two compounds which predominate in T2 deoxyribonucleic acid, deoxy-5-hydroxymethylcytidine triphosphate and deoxy-5-monoglucosyl hydroxymethylcytidine triphosphate. These preparations required an expedient, large scale preparation of the deoxyribonucleic acid of T2 phage, isolation of the required nucleoside monophosphates, and synthesis of the di- and triphosphates by the procedure of Clark et al. (4). In the course of this investigation, a convenient method was discovered for the preparation of crystalline deoxy-5-hydroxymethylcytidine monophosphate. This paper describes these procedures in detail.