rec-A protein-promoted recombination reaction consists of two independent processes, homologous matching and processive unwinding. A study involving an anti-rec-A protein-monoclonal IgG.
recA protein promotes the formation and processing of joint molecules of homologous double-stranded DNA and single-stranded DNA. We studied the effects of an anti-recA protein monoclonal IgG (ARM193) on two processes carried out by the recA protein. The homologous matching, i.e. pairing of double-stranded DNA and single-stranded DNA by forming intermolecular base-pairing at homologous regions was found to occur even in the presence of an excess amount of antibody ARM193. On the other hand, processive unwinding, i.e. the propagation of the unwinding of double-stranded DNA through a processive reaction of recA protein, which occurs even in the absence of single-stranded DNA, was found to be very sensitive to the inhibition by antibody ARM193. Therefore, we conclude that homologous matching and processive unwinding are independent of each other. Analysis of the effect of antibody ARM193 on the various activities of recA protein suggests that the entire reaction of the formation of joint molecules and their processing can be rationalized in terms of these two underlying processes, homologous matching and processive unwinding. This analysis also suggests that homologous matching seems to require only the binding itself of active units of recA protein to single-stranded DNA but not necessarily either the cooperativity of the protein or unwinding.
D000818:Animals; D000911:Antibodies, Monoclonal; D004277:DNA, Single-Stranded / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D007074:Immunoglobulin G / Q000276:immunology*; D051379:Mice; D011061:Poly A / Q000378:metabolism; D011938:Rec A Recombinases / Q000494:pharmacology*; D011995:Recombination, Genetic
D000911:Antibodies, Monoclonal; D004277:DNA, Single-Stranded; D007074:Immunoglobulin G; D011061:Poly A; C015465:poly(dA); D011938:Rec A Recombinases