Characterization of 2'(3')-trinitrophenyl-ATP as an inhibitor of ATP-dependent initiation complex formation between the DNA polymerase III holoenzyme and primed DNA

J Biol Chem. 1987 Mar 25;262(9):4190-4.


We have identified 2'(3')-trinitrophenyl-ATP to be an inhibitor of the ATP-dependent initiation complex formation reaction between the Escherichia coli DNA polymerase III holoenzyme and primed DNA. The inhibitor is specific for the initiation stage; once initiation complexes are formed the subsequent elongation reaction is unaffected. Three ATP-dependent DNA polymerase III holoenzyme reactions can be independently assayed: the ATP-dependent formation of initiation complexes, ATP binding, and the primed DNA-dependent hydrolysis of ATP. Trinitrophenyl ATP inhibits all three reactions to a similar extent with an apparent Ki between 6 and 15 microM in the presence of 5 microM ATP. This suggests all of these reactions are related and that they proceed through a common ATP-binding site. We include an improved purification of the DNA polymerase III holoenzyme in this report.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / analogs & derivatives*
  • Adenosine Triphosphate / metabolism
  • Adenosine Triphosphate / pharmacology*
  • Binding Sites
  • DNA / metabolism*
  • DNA Polymerase III / antagonists & inhibitors*
  • DNA Polymerase III / metabolism
  • Escherichia coli / enzymology
  • Hydrolysis
  • Nucleic Acid Synthesis Inhibitors*


  • 2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate
  • Nucleic Acid Synthesis Inhibitors
  • Adenosine Triphosphate
  • DNA
  • DNA Polymerase III