Interaction of recA protein with left-handed Z-DNA.
The ability of recA protein to interact with a Z-DNA polymer, Br-poly(dG-dC), or M13 bacteriophage single-stranded DNA was investigated. RecA protein binds more avidly to Z-DNA than to single-stranded DNA in the absence of a nucleotide cofactor. This binding pattern changes in the presence of adenosine 5'-(gamma-thio)triphosphate (ATP[S]), however, such that the binding to Z-DNA decreases while binding to single-stranded DNA increases roughly 2-fold. When present together, the two forms of DNA compete with each other in the presence of ATP[S]. Experiments involving recA protein binding to recombinant plasmids showed neither a preferential binding of recA protein to the plasmid containing Z-DNA nor a similar effect of ATP[S] to that observed with the Z-DNA polymer. In contrast, maximal binding was obtained with a plasmid (linear or supercoiled) containing a polypurine.polypyrimidine insert, thus suggesting that recA protein displays sequence preferences in its interaction with DNA. The results of the present study provide no evidence that recA protein specifically interacts with or stabilizes the Z-DNA insert of a recombinant plasmid in the left-handed conformation.
Krishna
P
P
Department of Medical Biochemistry, University of Calgary, Alberta, Canada.
Morgan
A R
AR
van de Sande
J H
JH
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D001435:Bacteriophages / Q000235:genetics; D001667:Binding, Competitive; D003432:Cross-Linking Reagents; D004247:DNA / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D004926:Escherichia coli / Q000032:analysis; D009690:Nucleic Acid Conformation; D010957:Plasmids; D011089:Polydeoxyribonucleotides / Q000378:metabolism; D011938:Rec A Recombinases / Q000378:metabolism*
D003432:Cross-Linking Reagents; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D011089:Polydeoxyribonucleotides; C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; D004247:DNA; C043078:poly(dG-Br(5)dC).poly(dG-Br(5)dC); D011938:Rec A Recombinases