Partial purification and characterization of a recombinase from human cells.
We describe the partial purification and characterization of a human recombinase activity from RPMI 1788 B lymphoblasts. Stoichiometric amounts of recombinase carry out a strand transfer reaction between linear duplex DNA and homologous circular single-strand DNA. The product of strand transfer by the recombinase is a joint molecule composed of a single-strand circle joined to one end of the linear duplex molecule by a region of DNA heteroduplex at least 150 bp long. Formation of DNA heteroduplexes is accompanied by strand displacement. Strand invasion initiates at the ends of the linear duplex. Finally, strand displacement by human recombinase exhibits polarity and proceeds in a 3' to 5' direction. This is the first demonstration of a strand transfer activity from a high eukaryote. We discuss similarities between our recombinase and the RecA and rec1 recombination proteins from E. coli and Ustilago maydis, respectively.
D000255:Adenosine Triphosphate / Q000502:physiology; D001402:B-Lymphocytes / Q000201:enzymology*; D002460:Cell Line; D004247:DNA / Q000096:biosynthesis; D004254:DNA Nucleotidyltransferases / Q000302:isolation & purification* / Q000494:pharmacology; D004270:DNA, Circular / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D043211:Exodeoxyribonuclease V; D005656:Fungal Proteins / Q000494:pharmacology; D006801:Humans; D008854:Microscopy, Electron; D009693:Nucleic Acid Hybridization; D011938:Rec A Recombinases / Q000494:pharmacology; D011995:Recombination, Genetic / Q000187:drug effects*; D012689:Sequence Homology, Nucleic Acid
D004270:DNA, Circular; D004277:DNA, Single-Stranded; D005656:Fungal Proteins; D000255:Adenosine Triphosphate; D004247:DNA; D004254:DNA Nucleotidyltransferases; D011938:Rec A Recombinases; D043211:Exodeoxyribonuclease V