Stereochemical course of nucleotidyl transfer catalyzed by bacteriophage T7 induced DNA polymerase

Biochemistry. 1982 May 11;21(10):2570-2. doi: 10.1021/bi00539a042.


The bacteriophage T7 induced DNA polymerase, consisting of the phage specified gene 5 protein associated with Escherichia coli thioredoxin, catalyzes the copolymerization of SP-dATP alpha S with dTTP, producing the alternating of polymer poly[dTs-A)] by a mechanism involving inversion of configuration at P alpha. Degradation of poly[d(5s-A)] by the nucleolytic action of E. coli DNA polymerase produced the dinucleotide pdTps-dA, whose configuration at the phosphorothioate diester was assigned as R by comparison of the phosphorus-31 nuclear magnetic resonance chemical shift (55.0 ppm downfield from H3PO4) with that of an authentic sample. Further degradation by alkaline phosphatase to Rp-dTps-dA (55.6 ppm downfield from H3PO4) confirmed the configuration. The stereochemistry provides no evidence of a double displacement mechanism.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • DNA-Directed DNA Polymerase / metabolism*
  • Deoxyadenine Nucleotides
  • Deoxyribonucleotides
  • Models, Chemical
  • Phosphoproteins
  • Stereoisomerism
  • T-Phages / enzymology*
  • Thionucleotides
  • Thioredoxins / analogs & derivatives


  • Deoxyadenine Nucleotides
  • Deoxyribonucleotides
  • Phosphoproteins
  • Thionucleotides
  • Thioredoxins
  • 2'-deoxyadenosine 5'-O-(1-thiotriphosphate)
  • bacteriophage T7 induced DNA polymerase
  • DNA-Directed DNA Polymerase