title: RNA-DNA hybridization promoted by E. coli RecA protein.
abstract: RecA protein of E. coli plays a central regulatory role that is induced by damage to DNA and results in the inactivation of LexA repressor. In vitro, RecA protein binds preferentially to single-stranded DNA to form a nucleoprotein filament that can recognize homology in naked duplex DNA and promote extensive strand exchange. Although RecA protein shows little tendency at neutral pH to bind to RNA, we found that it nonetheless catalyzed at 37 degrees C the hybridization of complementary RNA and single-stranded DNA sequences. Hybrids made by RecA protein at 37 degrees C appeared indistinguishable from ones prepared by thermal annealing. RNA-DNA hybridization by RecA protein at neutral pH required, as does RecA-promoted homologous pairing, optimal conditions for the formation of RecA nucleoprotein filaments. The cosedimentation of RNA with those filaments further paralleled observations made on the formation of networks of nucleoprotein filaments with double-stranded DNA, an instrumental intermediate in homologous pairing in vitro. These similarities with the pairing reaction support the view that RecA protein acts specifically in the hybridization reaction.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06510.
authors.forename: D P
authors.initials: DP
authors.lastname: Kirkpatrick
authors.forename: B J
authors.initials: BJ
authors.lastname: Rao
authors.forename: C M
authors.initials: CM
authors.lastname: Radding
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004586:Electrophoresis; D004926:Escherichia coli / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000378:metabolism; D009693:Nucleic Acid Hybridization / Q000502:physiology*; D012313:RNA / Q000378:metabolism*; D011938:Rec A Recombinases / Q000378:metabolism*; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D004277:DNA, Single-Stranded; D012313:RNA; D011938:Rec A Recombinases; D008274:Magnesium