The herpes simplex virus type-1 single-strand DNA-binding protein (ICP8) promotes strand invasion.
ICP8, the herpes simplex virus type-1 single-strand DNA-binding protein, was recently shown to promote strand exchange in conjunction with the viral replicative helicase (Nimonkar, A. V., and Boehmer, P. E. (2002) J. Biol. Chem. 277, 15182-15189). Here we show that ICP8 also catalyzes strand invasion in an ATP-independent manner. Thus, ICP8 promotes the assimilation of a single-stranded donor molecule into a homologous plasmid, resulting in the formation of a displacement loop. Invasion of a homologous duplex by single-stranded DNA requires homology at either 3' or 5' end of the invading strand. The reaction is dependent on the free energy of supercoiling and alters the topology of the acceptor plasmid. Hence, strand invasion products formed by ICP8 are resistant to the action of restriction endonucleases that cleave outside of the area of pairing. The ability to catalyze strand invasion is a novel activity of ICP8 and the first demonstration of a eukaryotic viral single-strand DNA-binding protein to promote this reaction. In this regard ICP8 is functionally similar to the prototypical prokaryotic recombinase RecA and its eukaryotic homologs. This strand invasion activity of ICP8 coupled with DNA synthesis may explain the high prevalence of branched DNA structures during viral replication.
Nimonkar
Amitabh V
AV
Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Miami, Florida 33101-6129, USA.
Boehmer
Paul E
PE
D000255:Adenosine Triphosphate / Q000378:metabolism; D002384:Catalysis; D004268:DNA-Binding Proteins; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D004720:Endonucleases / Q000378:metabolism; D004996:Ethidium / Q000494:pharmacology; D009696:Nucleic Acids / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism; D011938:Rec A Recombinases / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism
D004268:DNA-Binding Proteins; C045462:ICP8 protein, Simplexvirus; D009696:Nucleic Acids; D014764:Viral Proteins; D000255:Adenosine Triphosphate; D011938:Rec A Recombinases; D004720:Endonucleases; D004996:Ethidium