Synthesis of N‐Acetylated Deoxyribonucleoside 5′‐Triphosphates and their Utilization in Enzymatic Formation of Single‐Stranded Polydeoxyribonucleotides
Note. Oligo‐ and polydeoxynucleotides are designated according to the IUPAC‐IUB Combined Commission on Biochemical Nomenclature (CBN); overlining of a subscript means that the number is the average of a distribution; the superscripts Ac or Ac2 following a capital letter show that the nucleotide is acetylated or diacetylated; monomer units in a polymer are referred to by dA, dGAc, etc.; labeling in compunds is sparingly designated, usually just at the start of an experiment; the mono‐μmole is a micromole of monomer units contained in a polymer.
Abstract
The N‐acetylated deoxyadenosine and deoxyguanosine 5′‐triphosphates (dAAcTP and dGAcTP) have been synthesized and found competent as substrates for enzymatic chain lengthening of polydeoxyribonucleotides without concomitant loss of the acetyl group. The enzyme used was terminal deoxyribonucleotidyltransferase. The addition of dGAc units proceeds without aggregation that self‐limits addition of nonacetylated dG units. The resulting polymer 3′‐ended with dGAc units is an efficient initiator for subsequent chain lengthening. Hydrogen bonding is not destroyed by N‐acetylation of the dA‐dT Watson‐Crick pair, although it is considerably weakened. As a practical result of this, a polymer 3′‐ended with dAAc units is a much more efficient initiator for enzymatic chain lengthening by dT units than is a corresponding polymer with dA units. Venom phosphodiesterase does not hydrolyze a polymer 3′‐ended with dAAc units.
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