DNA-strand exchange promoted by RecA protein in the absence of ATP: implications for the mechanism of energy transduction in protein-promoted nucleic acid transactions

Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3478-82. doi: 10.1073/pnas.92.8.3478.

Abstract

DNA-strand exchange promoted by Escherichia coli RecA protein normally requires the presence of ATP and is accompanied by ATP hydrolysis, thereby implying a need for ATP hydrolysis. Previously, ATP hydrolysis was shown not to be required; here we demonstrate furthermore that a nucleoside triphosphate cofactor is not required for DNA-strand exchange. A gratuitous allosteric effector consisting of the noncovalent complex of ADP and aluminum fluoride, ADP.AIF4-, can both induce the high-affinity DNA-binding state of RecA protein and support the homologous pairing and exchange of up to 800-900 bp of DNA. These results demonstrate that induction of the functionally active, high-affinity DNA-binding state of RecA protein is needed for RecA protein-promoted DNA-strand exchange and that there is no requirement for a high-energy nucleotide cofactor for the exchange of DNA strands. Consequently, the free energy needed to activate the DNA substrates for DNA-strand exchange is not derived from ATP hydrolysis. Instead, the needed free energy is derived from ligand binding and is transduced to the DNA via the associated ligand-induced structural transitions of the RecA protein-DNA complex; ATP hydrolysis simply destroys the effector ligand. This concept has general applicability to the mechanism of energy transduction by proteins.

Publication types

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

MeSH terms

  • Adenosine Diphosphate / metabolism
  • Adenosine Diphosphate / pharmacology
  • Adenosine Triphosphate / analogs & derivatives*
  • Adenosine Triphosphate / metabolism
  • Aluminum Compounds / metabolism
  • Aluminum Compounds / pharmacology
  • DNA-Binding Proteins / drug effects
  • DNA-Binding Proteins / metabolism*
  • Energy Metabolism*
  • Fluorides / metabolism
  • Fluorides / pharmacology
  • Models, Genetic
  • Nucleic Acid Heteroduplexes / metabolism
  • Protein Binding / drug effects
  • Rec A Recombinases / drug effects
  • Rec A Recombinases / metabolism*
  • Recombination, Genetic / physiology*
  • Sodium Chloride / pharmacology

Substances

  • Aluminum Compounds
  • DNA-Binding Proteins
  • Nucleic Acid Heteroduplexes
  • tetrafluoroaluminate
  • Sodium Chloride
  • Adenosine Diphosphate
  • Adenosine Triphosphate
  • Rec A Recombinases
  • Fluorides