Thermochemical and kinetic evidence for nucleotide-sequence-dependent RecA-DNA interactions.
RecA catalyses homologous recombination in Escherichia coli by promoting pairing of homologous DNA molecules after formation of a helical nucleoprotein filament with single-stranded DNA. The primary reaction of RecA with DNA is generally assumed to be unspecific. We show here, by direct measurement of the interaction enthalpy by means of isothermal titration calorimetry, that the polymerisation of RecA on single-stranded DNA depends on the DNA sequence, with a high exothermic preference for thymine bases. This enthalpic sequence preference of thymines by RecA correlates with faster binding kinetics of RecA to thymine DNA. Furthermore, the enthalpy of interaction between the RecA x DNA filament and a second DNA strand is large only when the added DNA is complementary to the bound DNA in RecA. This result suggests a possibility for a rapid search mechanism by RecA x DNA filaments for homologous DNA molecules.
Wittung
P
P
Department of Physical Chemistry, Chalmers University of Technology, Gothenburg, Sweden.
Ellouze
C
C
Maraboeuf
F
F
Takahashi
M
M
Nordèn
B
B
D001665:Binding Sites / Q000235:genetics; D002151:Calorimetry; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011938:Rec A Recombinases / Q000378:metabolism*; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA
D004277:DNA, Single-Stranded; D011938:Rec A Recombinases